High-throughput screening of small molecules in miniaturized mammalian cell-based assays involving post-translational modifications. (1/58879)

BACKGROUND: Fully adapting a forward genetic approach to mammalian systems requires efficient methods to alter systematically gene products without prior knowledge of gene sequences, while allowing for the subsequent characterization of these alterations. Ideally, these methods would also allow function to be altered in a temporally controlled manner. RESULTS: We report the development of a miniaturized cell-based assay format that enables a genetic-like approach to understanding cellular pathways in mammalian systems using small molecules, rather than mutations, as the source of gene-product alterations. This whole-cell immunodetection assay can sensitively detect changes in specific cellular macromolecules in high-density arrays of mammalian cells. Furthermore, it is compatible with screening large numbers of small molecules in nanoliter to microliter culture volumes. We refer to this assay format as a 'cytoblot', and demonstrate the use of cytoblotting to monitor biosynthetic processes such as DNA synthesis, and post-translational processes such as acetylation and phosphorylation. Finally, we demonstrate the applicability of these assays to natural-product screening through the identification of marine sponge extracts exhibiting genotype-specific inhibition of 5-bromodeoxyuridine incorporation and suppression of the anti-proliferative effect of rapamycin. CONCLUSIONS: We show that cytoblots can be used for high-throughput screening of small molecules in cell-based assays. Together with small-molecule libraries, the cytoblot assay can be used to perform chemical genetic screens analogous to those used in classical genetics and thus should be applicable to understanding a wide variety of cellular processes, especially those involving post-transitional modifications.  (+info)

Retinoic acid, but not arsenic trioxide, degrades the PLZF/RARalpha fusion protein, without inducing terminal differentiation or apoptosis, in a RA-therapy resistant t(11;17)(q23;q21) APL patient. (2/58879)

Primary blasts of a t(11;17)(q23;q21) acute promyelocytic leukaemia (APL) patient were analysed with respect to retinoic acid (RA) and arsenic trioxide (As2O3) sensitivity as well as PLZF/RARalpha status. Although RA induced partial monocytic differentiation ex vivo, but not in vivo, As203 failed to induce apoptosis in culture, contrasting with t(15;17) APL and arguing against the clinical use of As203 in t(11;17)(q23;q21) APL. Prior to cell culture, PLZF/RARalpha was found to exactly co-localize with PML onto PML nuclear bodies. However upon cell culture, it quickly shifted towards microspeckles, its localization found in transfection experiments. Arsenic trioxide, known to induce aggregation of PML nuclear bodies, left the microspeckled PLZF/RARalpha localization completely unaffected. RA treatment led to PLZF/RARalpha degradation. However, this complete PLZF/RARalpha degradation was not accompanied by differentiation or apoptosis, which could suggest a contribution of the reciprocal RARalpha/PLZF fusion product in leukaemogenesis or the existence of irreversible changes induced by the chimera.  (+info)

Complete nucleotide sequence of the 27-kilobase virulence related locus (vrl) of Dichelobacter nodosus: evidence for extrachromosomal origin. (3/58879)

The vrl locus is preferentially associated with virulent isolates of the ovine footrot pathogen, Dichelobacter nodosus. The complete nucleotide sequence of this 27.1-kb region has now been determined. The data reveal that the locus has a G+C content much higher than the rest of the D. nodosus chromosome and contains 22 open reading frames (ORFs) encoding products including a putative adenine-specific methylase, two potential DEAH ATP-dependent helicases, and two products with sequence similarity to a bacteriophage resistance system. These ORFs are all in the same orientation, and most are either overlapping or separated by only a few nucleotides, suggesting that they comprise an operon and are translationally coupled. Expression vector studies have led to the identification of proteins that correspond to many of these ORFs. These data, in combination with evidence of insertion of vrl into the 3' end of an ssrA gene, are consistent with the hypothesis that the vrl locus was derived from the insertion of a bacteriophage or plasmid into the D. nodosus genome.  (+info)

Relationship between UDP-glucose 4-epimerase activity and oligoglucose glycoforms in two strains of Neisseria meningitidis. (4/58879)

Sodium dodecyl sulfate-polyacrylamide gel analysis of lipooligosaccharide (LOS) from Neisseria meningitidis has demonstrated considerable microheterogeneity in the variable region of LOS due to the presence of novel glycoforms. As a step toward understanding the basis for the expression of these novel glycoforms, we have examined the LOS structures and UDP-glucose 4-epimerase (epimerase) activity levels in two strains (NMB and MA-1) and their respective galE mutants. Strain NMB was found to have low epimerase activity and to contain multiple glycoforms, some of which appear to contain only glucose sugars. The galE mutant had only the oligoglucose glycoforms. Strain MA-1 had higher epimerase activity at both log and stationary phases (2- and 12.5-fold, respectively) and one glycoform with a putative lactosyl structure. Strain MA-1 galE had two glycoforms that contained one or two glucose residues. To understand the molecular basis for the different epimerase activities, we examined the predicted amino acid sequences of the respective galE open reading frames and determined the relative amounts of GalE protein. We found no significant differences between the predicted amino acid sequence of the GalE protein in NMB and that in MA-1. We observed no significant differences in the level of GalE protein between MA-1 and NMB at exponential or stationary phase. We also observed an 8.2-fold drop in epimerase activity in NMB between the log and stationary phases that was not due to the GalE protein level or low glucose levels.  (+info)

Characterization of human bactericidal antibodies to Bordetella pertussis. (5/58879)

The Bordetella pertussis BrkA protein protects against the bactericidal activity of complement and antibody; however, some individuals mount an immune response that overcomes this bacterial defense. To further characterize this process, the bactericidal activities of sera from 13 adults with different modes of exposure to B. pertussis (infected as adults, occupational exposure, immunized with an acellular vaccine, or no identified exposure) against a wild-type strain and a BrkA complement-sensitive mutant were evaluated. All of the sera killed the BrkA mutant, suggesting past exposure to B. pertussis or cross-reactive organisms. Several samples had no or minimal activity against the wild type. All of the sera collected from the infected and occupationally exposed individuals but not all of the sera from vaccinated individuals had bactericidal activity against the wild-type strain, suggesting that some types of exposure can induce an immune response that can overcome the BrkA resistance mechanism. Adsorbing serum with the wild-type strain removed the bactericidal antibodies; however, adsorbing the serum with a lipopolysaccharide (LPS) mutant or an avirulent (bvg mutant) strain did not always result in loss of bactericidal activity, suggesting that antibodies to either LPS or bvg-regulated proteins could be bactericidal. All the samples, including those that lacked bactericidal activity, contained antibodies that recognized the LPS of B. pertussis. Bactericidal activity correlated best with the presence of the immunoglobulin G3 (IgG3) antibodies to LPS, the IgG subtype that is most effective at fixing complement.  (+info)

Characterization of Moraxella (Branhamella) catarrhalis lbpB, lbpA, and lactoferrin receptor orf3 isogenic mutants. (6/58879)

Pathogenic members of the family Neisseriaceae produce specific receptors to acquire iron from their host's lactoferrin and transferrin. Recently, putative Moraxella catarrhalis lactoferrin receptor genes and a third open reading frame (lbpB, lbpA, and orf3) were cloned and sequenced. We describe the preliminary characterization of isogenic mutants deficient in LbpB, LbpA, or Orf3 protein.  (+info)

Mechanisms related to [18F]fluorodeoxyglucose uptake of human colon cancers transplanted in nude mice. (7/58879)

[18F]Fluorodeoxyglucose ([18F]FDG), a glucose analogue, has been widely used for tumor imaging. To investigate the mechanisms related to [18F]FDG uptake by tumors, an experiment involving nude mice was performed. METHODS: Human colon cancer cell lines SNU-C2A, SNU-C4 and SNU-C5 were transplanted to nude mice. Using immunohistochemical staining and Western blot, the expression of glucose transporter (Glut) isoforms (Glut-1 through -5) in xenografted tumors was analyzed. For the analysis of messenger ribonucleic acid (mRNA) expression, reverse-transcription polymerase chain reaction and Northern blot were used and the enzyme activity of hexokinase in cancer tissues was measured by continuous spectrophotometric rate determination. RESULTS: [18F]FDG uptake in SNU-C4 and SNU-C5 cells was higher than in normal colon cells. Among these cells and xenografted tumors, SNU-C5 showed the highest level of [18F]FDG uptake, followed by SNU-C4 and SNU-C2A. An immunostaining experiment showed intense staining of Glut-1 in SNU-C5 tumors but somewhat faint staining in SNU-C4. SNU-C5 tumors also showed positive staining with Glut-3, although this was not the case with SNU-C2A and SNU-C4. Western blot analysis showed the expression of Glut-1 and Glut-3 in all tumors. Experiments involving Northern blot analysis and reverse-transcription polymerase chain reaction confirmed the overexpression of Glut-1 mRNA in all tumors, with the highest level in SNU-C5. The level of Glut-3 mRNA was also elevated in SNU-C5 tumors but not in SNU-C2A and SNU-C4. The enzyme activity of hexokinase did not vary among different tumors. CONCLUSION: Gluts, especially Glut-1, are responsible for [18F]FDG uptake in a nude mouse model of colon cancer rather than hexokinase activity. Increased numbers of glucose transporters at the plasma membrane of cancer cells is attributed to an increased level of transcripts of glucose transporter genes and may be a cause of increased [18F]FDG uptake, at least in colon cancer tumors.  (+info)

Cloning of the peroxiredoxin gene family in rats and characterization of the fourth member. (8/58879)

Peroxiredoxin (PRx) exhibits thioredoxin-dependent peroxidase activity and constitutes a family of proteins. Four members of genes from rat tissues were isolated by PCR using degenerated primers based on the sequences which encode a pair of highly conserved Cys-containing domains, and were then cloned to full-length cDNAs. These included two genes which have previously been isolated in rats, PRx I and PRx II, and two rat homologues of PRx III and PRx IV. We showed, for the first time, the simultaneous expression of all four genes in various rat tissues by Northern blotting. Since a discrepancy exists regarding cellular distribution, we further characterized PRx IV by expressing it in COS-1 cells. This clearly demonstrates that PRx IV is a secretory form and functions within the extracellular space.  (+info)

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"Western Blotting Inaccuracies with Unverified Antibodies: Need for a Western Blotting Minimal Reporting Standard (WBMRS)". PLOS ... Normalization of Western blot data is an analytical step that is performed to compare the relative abundance of a specific ... 2015-01-01). "Western Blotting Using In-Gel Protein Labeling as a Normalization Control: Stain-Free Technology". In Posch, ... When using fluorescent antibodies to image proteins in western blots, normalization requires that the user define the upper and ...
The far-western blot, or far-western blotting, is a molecular biological method based on the technique of western blot to ... blotting' step. In a western blot, specific proteins are then identified using an antibody probe. Far-western blot employs non- ... Whereas western blot uses an antibody probe to detect a protein of interest, far-western blot uses a non-antibody probe which ... Thus, whereas western blotting is used for the detection of certain proteins, far-western blotting is employed to detect ...
Western blot Northwestern blot Far-eastern blot Blot Freeze, HH (1993). "Preparation and analysis of glycoconjugates". Current ... Since this is essentially a western blot, the charge reversal was used to dub this method an eastern blot. (2005) Eastern blot ... distinguishing them from a standard far-western blot. In principle, eastern blotting is similar to lectin blotting (i.e., ... Thus, eastern blot can be considered an extension of the biochemical technique of western blot. Multiple techniques have been ...
Southern blot for DNA northern blot for RNA reverse northern blot for RNA western blot for proteins far-western blot for ... making the Far-Western blot a specific case of the Western blot. A southwestern blot is based on Southern blot and is used to ... Similar to a western blot, the far-western blot uses protein-protein interactions to detect the presence of a specific protein ... A dot blot is a special case of any of the above blots where the analyte is added directly to the blotting matrix (and appears ...
Southern blot Northern blot Western blot Southwestern blot "Test Blots, Slot Blots & Dot Blots - Immunodetection , Bio-Rad". ... Performing a dot blot is similar in idea to performing a western blot, with the advantage of faster speed and lower cost. Dot ... or slot blot) is a technique in molecular biology used to detect proteins. It represents a simplification of the western blot ... A general dot blot protocol involves spotting 1-2 microliters of a samples onto a nitrocellulose or PVDF membrane and letting ...
The name originates from a combination of ideas underlying Southern blotting and Western blotting techniques of which they ... Lelong, Jean-Claude; Prevost, Grégoire; Lee, Kyong il; Crepin, Michel (June 1989). "South Western blot mapping: A procedure for ... Thereafter southwestern blotting begins to vary with regards to procedure as since the first blotting's, many more have been ... Southwestern+Blot at the U.S. National Library of Medicine Medical Subject Headings (MeSH) Bowen, B; Steinberg, J; Laemmli, U K ...
Kurien BT, Scofield RH (April 2006). "Western blotting". Methods. 38 (4): 283-93. doi:10.1016/j.ymeth.2005.11.007. PMID ... in Western blot analyses to identify proteins separated by electrophoresis, and in immunohistochemistry or immunofluorescence ... Western blot, immunodiffusion, immunoelectrophoresis, and magnetic immunoassay. Antibodies raised against human chorionic ... antibody Optimer Ligand Secondary antibodies Single-domain antibody Slope spectroscopy Synthetic antibody Western blot ...
... western blot for protein, or for DNA extraction). Most lysis buffers contain buffering salts (e.g. Tris-HCl) and ionic salts (e ... Western Blotting. Methods in Molecular Biology. Vol. 1312. Springer New York. pp. 49-60. doi:10.1007/978-1-4939-2694-7_8. ISBN ...
A western blot is a technique by which specific proteins can be detected from a mixture of proteins. Western blots can be used ... "Western blot , Learn Science at Scitable". www.nature.com. Retrieved 2021-11-25. Kurien BT, Scofield RH (April 2006). "Western ... Using western blotting techniques allows not only detection but also quantitative analysis. Analogous methods to western ... In western blotting, proteins are first separated by size, in a thin gel sandwiched between two glass plates in a technique ...
and Western blotting Löf, Liza; Arngården, Linda; Ebai, Tonge; Landegren, Ulf; Söderberg, Ola; Kamali-Moghaddam, Masood (2017 ... November 2011). "Western blotting via proximity ligation for high performance protein analysis". Molecular & Cellular ...
Western blotting is a process by which proteins separated in the acrylamide gel are electrophoretically transferred to a stable ... Western blot). After staining, different species biomolecules appear as distinct bands within the gel. It is common to run ... Agarose gel electrophoresis Capillary electrophoresis DNA electrophoresis Eastern blotting Electroblotting Fast parallel ... focusing Isotachophoresis Native gel electrophoresis Northern blotting Protein electrophoresis QPNC-PAGE Southern blotting Two ...
Other blotting methods (i.e., western blot, northern blot, eastern blot, southwestern blot) that employ similar principles, but ... acids Restriction fragment Genetic fingerprint Northern blot Western blot Eastern blot Southwestern blot Northwestern blot " ... Burnette, W. Neal (April 1981). "Western Blotting: Electrophoretic Transfer of Proteins from Sodium Dodecyl Sulfate- ... The names for other blotting methods may follow this convention, by analogy. Southern invented Southern blot after combining ...
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Western blot Eastern blot Northwestern blot Far-eastern blot Far-western blot Differential display Gilbert, S. F. (2000) ... Northern blotting takes its name from its similarity to the first blotting technique, the Southern blot, named for biologist ... The northern blot, or RNA blot, is a technique used in molecular biology research to study gene expression by detection of RNA ... The major difference is that RNA, rather than DNA, is analyzed in the northern blot. A general blotting procedure starts with ...
Development of Western blotting technique to detect proteins. Publication of two protocols for plant transgenesis, which were ...
northern blotting A blotting method in molecular biology used to detect RNA in a sample. Compare Southern blotting, western ... Top A B C D E F G H I J K L M N O P Q R S T U V W X Y Z See also References External links Warburg effect western blotting ... Southern blotting A molecular biology method used for detecting a specific sequence in DNA samples. The technique combines ... blotting, and eastern blotting. nRNA See nuclear RNA. nuclear cage nuclear envelope A sub-cellular barrier consisting of two ...
Alternatively the proteins may react with ligands like lectins giving rise to the term affinity blotting. Western blotting SDS- ... Protein (Western) Blotting - Introduction to Antibodies Membrane Transfer Detailed electroblotting to PVDF procedure - Protein ... Following electrophoresis, a standard tank or semi-dry blotting transfer system is set up. A stack is put together in the ... Because the proteins may retain or regain part of their structure during blotting they may react with specific antibodies ...
Low-to-mid-plex techniques: Reporter gene Northern blot Western blot Fluorescent in situ hybridization Reverse transcription ... One of the most commonly used methods is to perform a Western blot against the protein of interest. This gives information on ... The detection process is very similar to that of a Western blot, but by avoiding the gel steps more accurate quantification can ... Moritz, CP (10 February 2020). "40 years Western blotting: A scientific birthday toast". Journal of Proteomics. 212: 103575. ...
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  • Other related techniques include dot blot analysis, quantitative dot blot, immunohistochemistry and immunocytochemistry, where antibodies are used to detect proteins in tissues and cells by immunostaining, and enzyme-linked immunosorbent assay (ELISA). (wikipedia.org)
  • With direct detection, primary antibodies are conjugated directly to an enzyme, like HRP , for chemiluminescent detection or to fluorochromes including Alexa Fluor® dyes, Janelia Fluor® dyes or DyLight® dyes for fluorescent Western blotting. (novusbio.com)
  • Both monoclonal and polyclonal antibodies are used for immunoprobing of antigens in western blotting. (bdbiosciences.com)
  • The tagged antibodies are detected directly (direct western blotting) or using secondary antibodies (indirect western blotting). (bdbiosciences.com)
  • When incubated with a blot on which HRP-conjugated antibodies (or other probes) are bound, a chemical reaction emits light at 425 nm which can be captured with x-ray film, CCD camera imaging devices and phosphorimagers that detect chemiluminescence. (thermofisher.com)
  • The chemiPRO is our complete solution, featuring a highly sensitive cooled CCD camera with high quantum efficiency, for detection of picogram level proteins, this system can image both conventional ECL style chemiluminescence blots and fluorescently tagged proteins of antibodies with the optional fluorescence module. (cleaverscientific.com)
  • The IgG -WB was done in house and the test was considered positive if the child had antibodies that recognized at least one band on IgG blots different from the mother 's or with greater intensity than the corresponding maternal band, during the first three months of life . (bvsalud.org)
  • The western blot (sometimes called the protein immunoblot), or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. (wikipedia.org)
  • The western blot is extensively used in biochemistry for the qualitative detection of single proteins and protein-modifications (such as post-translational modifications). (wikipedia.org)
  • Proteins from known HIV-infected cells are separated and blotted on a membrane as above. (wikipedia.org)
  • Western blotting-or immunoblotting-is a technique used to detect, characterize and quantitate proteins. (bdbiosciences.com)
  • The separated proteins are then transferred or blotted to a nitrocellulose or a PVDF membrane to immobilize the protein. (bdbiosciences.com)
  • Using western blotting, proteins can be characterized for several aspects, including their presence or absence, abundance, phosphorylation status or localization. (bdbiosciences.com)
  • In addition to these qualitative aspects, western blotting can also be used for relative quantitation of proteins based on the band intensity between experimental and control samples. (bdbiosciences.com)
  • Blotting is carried out to transfer proteins from the gel to a solid membrane. (ahdiagnostics.se)
  • The effectiveness of the transfer is heavily reliant on the gel acrylamide percentage, the molecular weight of electrophoresed proteins, and the selected blotting membrane. (ahdiagnostics.se)
  • The S canLater Western Blot Protein Ladder consists of seven biotinylated recombinant proteins as well as three prestained marker proteins. (moleculardevices.com)
  • Detection of the biotinylated proteins is enabled by adding Europium-labeled streptavidin to the blot at the secondary antibody step. (moleculardevices.com)
  • The ScanLater Western Blot Protein Ladder consists of seven biotinylated recombinant proteins at 10, 20, 40, 50, 80, 100 and 140 kD. (moleculardevices.com)
  • White bands are biotinylated protein used to estimate molecular weights of blotted proteins. (moleculardevices.com)
  • The migration distances of seven proteins in the ScanLater Western Blot Protein Ladder are plotted and fitted as a function of their respective molecular weights. (moleculardevices.com)
  • Using western blotting for verification can be a helpful tool, but correct interpretation of the results is crucial to avoid misidentification of cell lines with functional proteins as knockouts (and vice versa). (horizondiscovery.com)
  • Western blot analysis is a method widely used in the lab today because of its versatility in detecting and measuring specific target proteins in a sample. (labroots.com)
  • Western blotting is an important technique used to identify specific proteins from a complex mixture of proteins extracted from cells or body fluids. (lumigen.com)
  • Lumigen ® has a range of sensitive substrates for the detection of Western blotted proteins with sustained signal duration, reduced antibody usage and rapid onset of chemiluminescence. (lumigen.com)
  • Western blotting is a decades-old laboratory technique that is used to detect specific proteins from cell culture, tissue, or blood specimen. (championsoncology.com)
  • Since its initial description, western blotting has been used in all fields of biological and biomedical research because it is a straightforward and robust method for detecting specific proteins. (championsoncology.com)
  • Western blot allows researchers to determine and quantify the proteins existent within a cell, identifying specific proteins out of the mixture of proteins that are present in cell samples. (news-medical.net)
  • This application note describes two methods that allow you to complete the entire western blot workflow in less than 2-5 hours, depending on which time-saving steps you incorporate. (flinnsci.com)
  • The ScanLater Western Blot workflow follows standard gel loading and blotting methods up to the secondary antibody incubation step. (moleculardevices.com)
  • Introducing the chemiSOLO chemiluminescent imaging system, offering a high performance yet economical option for chemiluminescent blot imaging, without sacrificing a large space on your lab bench. (bioscience.co.uk)
  • Click here for information on chemiluminescent western blot detection can be found here . (thermofisher.com)
  • Fluorescent reagents are growing in popularity for western blotting because they offer increased time savings over chemiluminescent detection and reduced chemical waste compared to both chemiluminescent or chromogenic detection systems. (thermofisher.com)
  • While the detection limits are still not as low as chemiluminescent detection, fluorescent detection has the unique advantage of allowing multiple targets to be assayed for on the same blot at the same time without the need to strip and reprobe. (thermofisher.com)
  • This video will show you how to adjust the image appearance of your chemiluminescent Western blot with Image Studio 4.0 Software on the C-DiGit Blot Scanner. (licor.com)
  • These systems can handle fluorescence 1D and 2D gels as well as chemiluminescent blots. (syngene.com)
  • The Trans-Blot Turbo Transfer System is a high-performance western blotting transfer system designed to provide rapid transfers with high efficiency. (bio-rad.com)
  • Add the Trans-Blot Turbo Transfer System of your choice from the list below to your cart and enter promo code 25OFFTRANSBLOT in the promotion code field to receive your discount. (bio-rad.com)
  • However, in recent years the reproducibility and quality of western blotting data has been increasingly scrutinized. (labroots.com)
  • This webinar series will share tips and provide step-by-step guidance on how to improve the quality and reproducibility of data for both qualitative and quantitative western blotting, with attention on some of the newer guidelines. (labroots.com)
  • Champions Oncology provides Western Blotting protein detection assays, developed on the JESS ProteinSimple Analyzer, for higher data quality and reproducibility. (championsoncology.com)
  • Testing was also performed using an in-house anti-HEV Western Blot assay. (cdc.gov)
  • Further testing with the Wantai HEV-IgG ELISA assay and an in-house anti-HEV Western Blot assay was performed on a subset of 1803 of the 6000 samples. (cdc.gov)
  • Our cutting-edge Western Blotting assay consists of a capillary electrophoresis-based immunoassay for protein detection and quantitation that delivers sharper images and more reproducible results. (championsoncology.com)
  • To support your research needs, we offer several options to customize and integrate our Western Blotting assay. (championsoncology.com)
  • These days, immunohistochemical staining is just one of several established immunostaining techniques, including enzyme-linked immunosorbent assay, flow cytometry, immuno-electron microscopy, and Western blotting. (news-medical.net)
  • Title : Interpretation and Use of the Western Blot Assay for Serodiagnosis of Human Immunodeficiency Virus Type 1 Infections Corporate Authors(s) : Centers for Disease Control (U.S.) Published Date : July 21, 1989 Source : MMWR. (cdc.gov)
  • Researchers use fluorescent western blotting to quantify multiple targets on a single blot. (the-scientist.com)
  • The growing demand for multiplex western blotting has driven the development of many new fluorescent dyes (such as Alexa Fluor Plus conjugates). (thermofisher.com)
  • Use Hi LEDs for multiplexing RGB and IR fluorescent blots and gels, UV transilluminator for DNA/RNA gels, visible light converter screen for Coomassie blue and silver stained protein gels and blue converter screen for imaging 'safe' dyes. (syngene.com)
  • A semi-quantitative estimation of a protein can be derived from the size and color intensity of a protein band on the blot membrane. (wikipedia.org)
  • The blotting is performed by assembling filter papers, a membrane, and a gel into a sandwich. (ahdiagnostics.se)
  • Some forms of Lyme disease testing employ western blotting. (wikipedia.org)
  • Western blot technique uses three elements to achieve its task of separating a specific protein from a complex: separation by size, transfer of protein to a solid support, and marking target protein using a primary and secondary antibody to visualize. (wikipedia.org)
  • At the final step of Western blotting you will need to choose a detection method to visualize your results. (ahdiagnostics.se)
  • The detection reagents can be removed and the entire blot reprobed to visualize another protein or to optimize detection of the first protein. (thermofisher.com)
  • An optimize protocol will maximize your protein-of-interest's solubility and stability, which will allow you to better visualize your protein and reliably interpret your Western blot data. (bitesizebio.com)
  • In a previous post, I talked about immunostaining and touched on the principles and procedures of Western blotting, a technique that is widely used to quantify protein expression. (metheno.com)
  • In this webinar, Kelly Wolfe will discuss the dos and don'ts of protein electrophoresis and western blotting. (the-scientist.com)
  • Following sample prep, you need to do polyacrylamide gel electrophoresis (PAGE) and blotting. (ahdiagnostics.se)
  • The primary applications of this protein ladder include the estimation of protein molecular weight, the visualization of gel electrophoresis, and the evaluation of the transfer process from gel to blot. (moleculardevices.com)
  • These blue bands are seen on the electrophoresis gel and on the blot after transfer. (moleculardevices.com)
  • Dark bands are pre-stained protein used to monitor progress of protein electrophoresis, blot orientation, and assess transfer efficiency. (moleculardevices.com)
  • Whether you are looking to move out of the darkroom and invest in your first digital imaging system/Western blot imager or if you are looking to replace an old system, it is important to purchase the right equipment that will adapt with your research. (syngene.com)
  • Implementing a gel imaging system/Western blot imager eliminates the chemical waste associated with film processing. (syngene.com)
  • Date: October 14, 2021 Time: 9:00am (PDT), 12:00pm (EDT), 5:00pm GMT Western blot analysis is a method widely used in the lab today because of its versatility in detecting and measuring spe. (labroots.com)
  • Western Blot is a widely used, essential, and trusted technique for protein analysis of complex systems. (championsoncology.com)
  • The final immunostaining method is the Western blot method, a widely used technique that has firmly ingrained itself in the fields of cell and molecular biology. (news-medical.net)
  • In this webinar, R. Hal Scofield discusses hints and tips on how to generate clean and reproducible western blot data, even in tricky situations. (the-scientist.com)
  • In this webinar, Kelly Wolfe will discuss innovations that streamline western blotting workflows. (the-scientist.com)
  • In this webinar, Aldrin Gomes will reveal how to obtain reproducible, publication-ready western blot images. (the-scientist.com)
  • Our team of western blotting experts have put together this 3-part webinar series that is designed to help you avoid some of the pitfalls associated with western blotting and enable you to achieve publication-quality western blot results. (labroots.com)
  • Muscle histology, immunocytochemistry, and Western blot analysis can be done. (msdmanuals.com)
  • Knockout Validated Western Blot showing lysates of HeLa parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM Chloroquine. (novusbio.com)
  • No attempts have been made to qualify this product on blots made with HeLa cell, or any other antigen source. (calbiotech.com)
  • Western blotting versus ELISA: What are the differences? (bdbiosciences.com)
  • Like in western blotting, protein detection using an ELISA also uses antibody binding to antigens and could be accomplished with one antibody (direct ELISA) or using a primary and a secondary antibody (indirect ELISA). (bdbiosciences.com)
  • Crude antigenic preparations from Setaria equina were used in ELISA and Western blotting to examine cross-reaction with human sera from areas endemic for bancroftian filariasis. (who.int)
  • Des préparations d'antigènes bruts de Setaria equina ont été utilisées dans le cadre des méthodes ELISA et transfert Western afin d'étudier la réaction croisée avec des sérums humains en provenance de zones endémiques pour la filariose de Bancroft. (who.int)
  • So I'd confirm by Western that there is only one protein reacting with the antibody, but for quantitation I'd use dot blots. (bio.net)
  • Membranes are scanned on either the SpectraMax Paradigm or SpectraMax i3 Multi-Mode Microplate Reader with the ScanLater Western Blot Detection Cartridge,which utilizes time resolved fluorescence (TRF) detection of Europium (Eu). (moleculardevices.com)
  • Europium is resistant to photo-bleaching, so the western blot signal remains stable for weeks to months which enables repeated reading of membranes. (moleculardevices.com)
  • Blotting membranes are supplied in sheet form and as a 3m roll which can be cut to size. (cleaverscientific.com)
  • With a range of Vertical Gel Systems, Power Supplies, Blot Transfer Systems, Reagents and Chemicals and Gel Documentation and Analysis, we provide the complete solution for the Life Scientist. (cleaverscientific.com)
  • We retrospectively reviewed findings from clinical and laboratory examinations, IHC for dystrophin of muscle biopsy tissue, Western blot analysis, and multiplex polymerase chain reaction (PCR) examination of genomic DNA. (nih.gov)
  • In this study, we investigated the relationship between clinical characteristics, findings at immunohistochemistry (IHC) and Western blot, and the pattern of exon deletions in 24 male patients with dystrophinopathies. (nih.gov)
  • Representative western blotting analyses of brain insulin-induced signaling molecules. (novusbio.com)
  • Together with the advances in the digital imaging equipment these new fluorophores enable extremely powerful analyses in western blotting. (thermofisher.com)
  • Western blot and Wantai IgG anti-HEV testing was performed to determine correlation with the DSI and Kuniholm NIH in-house IgG anti-HEV assays. (cdc.gov)
  • At Champions, we provide Western Blotting protein detection assays, optimized on the Jess ProteinSimple Analyzer. (championsoncology.com)
  • A western blot can also be used as a confirmatory test for Hepatitis B infection and HSV-2 (Herpes Type 2) infection. (wikipedia.org)
  • Faster and more efficient than other western blotting transfer methods. (bio-rad.com)
  • The confirmatory HIV test employs a western blot to detect anti-HIV antibody in a human serum sample. (wikipedia.org)
  • The term "western blot" was given by W. Neal Burnette in 1981, although the method itself was independently invented in 1979 by Jaime Renart, Jakob Reiser, and George Stark at Stanford University, and by Harry Towbin, Theophil Staehelin, and Julian Gordon at the Friedrich Miescher Institute in Basel, Switzerland. (wikipedia.org)
  • 2013). 'Detection of cathepsin D in ewe's milk by Western Blotting method', Mljekarstvo , 63(1), str. (srce.hr)
  • Dolenčić Špehar I, Martinković F, Havranek J, Marinculić A, Tudor Kalit M, Kalit S. Detection of cathepsin D in ewe's milk by Western Blotting method. (srce.hr)
  • This paper shows the presence of cathepsin D and its forms in ewe's milk by modified Western Blotting method. (srce.hr)
  • In this blog we provide an overview of the western blot method and highlight its current applications in preclinical oncology research. (championsoncology.com)
  • Evaluation of the Western blotting method for the diagnosis of congenital toxoplasmosis. (bvsalud.org)
  • To evaluate the Western blotting method for the detection of IgG anti- Toxoplasma gondii (T. gondii) ( IgG -WB) in the serum of children with suspected congenital toxoplasmosis . (bvsalud.org)
  • GeneSys image capture software from Syngene makes imaging gels and western blots simple by using system protocols - just place your sample in the darkroom, position your sample and click the capture button for high quality images. (syngene.com)
  • With boxes that are correctly matched to common blot sizes, you can fit more blots on shakers/rockers while keeping reagent and antibody usage to an absolute minimum. (thomassci.com)
  • Western blot sample preparations, including lysis buffers, lysate from cell culture, lysate from tissues and determination of protein concentration. (abcam.com)
  • We have gel loading tips, sample loading buffers, protein ladders, transfer buffers, and much more for the protein separation and blotting steps of Western blotting. (ahdiagnostics.se)
  • Further applications of the western blot technique include its use by the World Anti-Doping Agency (WADA). (wikipedia.org)
  • Then maximise your space and budget with an imaging system/Western blot imaging system that is versatile and capable of imaging a wide range of applications in one system. (syngene.com)
  • On the basis of application, the western blotting market is divided into clinical diagnostics, agricultural applications, biomedical research, and some other related applications. (tmrresearchblog.com)
  • A robust, automated western blot system accelerates immunotherapy development. (the-scientist.com)
  • The ScanLater ™ Western Blot Protein Ladder is an essential component of the ScanLater ™ Western Blot Detection System. (moleculardevices.com)
  • They are also seen as dark bands when the ladder is read with the Scanlater Western Blot Detection System. (moleculardevices.com)
  • They are detected after incubation with Eu-streptavidin and read on the ScanLater Western Blot Detection System. (moleculardevices.com)
  • The ScanLater Western Blot Detection System, including the ScanLater Western Blot Protein Ladder, is a simple, sensitive, and stable platform that provides excellent protein analysis capability in a multi-mode microplate reader. (moleculardevices.com)
  • Whichever gel imaging system/Western blot imaging system you choose, you want a system that is intuitive and can be used by everyone without training. (syngene.com)
  • Select a Western blot imaging system that uses LEDs as they are environmentally friendly. (syngene.com)
  • Transfers with swirls, mystery protein splotches, loss of protein, or a general variability in transfer efficiency are common Western blot problems . (bitesizebio.com)
  • Further experiments with Interleukin-17 and Heat Shock Protein also exemplify the use of the ScanLater Western Blot Protein Ladder for estimating molecular weight. (moleculardevices.com)
  • Estimation of the molecular weight of IL-17 using the ScanLater Western Blot Protein Ladder. (moleculardevices.com)
  • Antibody against human SS-A (Ro), detectable bands at SS-A 60kD by Western blot immunoassay at 1:100 dilution using Hep-2 cellular extract blot. (calbiotech.com)
  • Our expert team can help you integrate Western Blotting analysis in your preclinical studies or clinical specialty testing for target validation, model and sample characterization, detection of post-translational modification, and much more. (championsoncology.com)
  • from preparation to separation and blotting as well as antibody incubation and, lastly, detection. (ahdiagnostics.se)
  • Western blotting is a simple and straightforward technique for protein characterization. (ahdiagnostics.se)
  • The western blot is routinely used for verification of protein production after cloning. (wikipedia.org)
  • Results of search for 'su:{Blotting, Western. (who.int)
  • Review tips to guide all stages of western blotting, from sample preparation to imaging and analysis. (the-scientist.com)
  • Users commonly look at the sensor resolution of the camera when evaluating gel documentation imaging systems/Western blot imagers. (syngene.com)
  • Western blot shows an increase in HSP70 that occurs when cells are exposed to temperatures that are higher than normal. (moleculardevices.com)