Proteins on a western blot blotting western. Medical search. Frequent questions

We now show that during early apoptosis limited membrane permeabilization occurs in blebs and apoptotic bodies, which allows release of proteins that may affect the proximal microenvironment before the catastrophic loss of membrane integrity during secondary necrosis. (nature.com)
Blebbing, apoptotic body formation and protein release during early apoptosis are dependent on ROCK and myosin ATPase activity to drive actomyosin contraction. (nature.com)
2 , 3 A key feature of apoptosis has been defined as the maintenance of an intact cellular membrane (detectable as exclusion of impermeable dyes such as propidium iodide, PI) throughout the apoptotic program to prevent intracellular protein release and subsequent immunological activation. (nature.com)
Our previous study has shown that CCAAT-enhancer binding protein β (C/EBPβ) is an important regulator in METH-induced neuronal autophagy and apoptosis. (cancerindex.org)
This proadifen-MTX synergism was also mediated by the inhibition of various cellular proteins engaged in apoptosis, including Mc-1, Bcl-xL, survivin and activation of procaspase-3. (spandidos-publications.com)

Here, using different experimental approaches, especially designer receptor exclusively activated by designer drug (DREADD) chemogenetic technology, we found that sustained activation of G q -protein signaling impairs the functionality of striatal neurons and we unveil the precise molecular mechanism underlying this process: a phospholipase C/Ca 2+ /proline-rich tyrosine kinase 2/cJun N -terminal kinase pathway. (jneurosci.org)
The expression of HMGB1, c-Jun N-terminal kinase (JNK), extracellular-signal-regulated kinase (ERK), and p38 mitogen-activated protein kinase (p38 MAPK) was assessed with immunofluorescence and western blot analysis. (molvis.org)
O 2 − release, protein kinase C (PKC) activity, and translocation of PKC-α and -βII and p47phox were increased in THP-1 cells (human monocytic cell line) under HG (15 mmol/l glucose) conditions, whereas AT supplementation inhibited these changes. (diabetesjournals.org)
The possible mechanisms by which hyperglycemia, in monocytes, can cause adverse effects is via the activation of diacylglycerol (DAG)-sensitive protein kinase C (PKC) ( 6 ). (diabetesjournals.org)
This gene is a member of the protein-tyrosine kinase oncogene family. (cancerindex.org)
The protein associates with the p85 subunit of phosphatidylinositol 3-kinase and interacts with the fyn-binding protein. (cancerindex.org)

When expressed as a recombinant protein in transfected cells, PV 3A cofractionates with endoplasmic reticulum markers ( 66 ), and its single transient expression can disrupt the secretory apparatus ( 23 ) and decrease major histocompatibility complex (MHC) class I expression ( 22 ). (asm.org)
There are no publications for TRIM50C Recombinant Protein (H00378108-P01). (novusbio.com)
Discover related pathways, diseases and genes to TRIM50C Recombinant Protein (H00378108-P01). (novusbio.com)

Total protein is ready for immediate use in Western blotting, immunoprecipitation, SDS-PAGE, isoelectric focusing gels and SDS-capillary electrophoresis, enzymatic activity analysis, gel shifting assay, protein-protein interaction, and tissue specific expression. (delos.info)

NS proteins are involved in crucial aspects of the viral cycle and pathogenesis, such as rearrangements of intracellular membranes required for endomembrane recruitment and the lysis of host cells ( 1 , 12 , 14 , 18 , 73 ). (asm.org)
in poliovirus (PV), the interaction between the RNA replication complex and intracellular membranes appears to be accomplished by proteins 3A and 2C, which have membrane-binding properties ( 11 , 60 ). (asm.org)
Using chimeric proteins in which CD4 is fused to the large intracellular loop of each of the AChR subunits we found that agrin induced clustering of only chimeras containing the β subunit loop. (jneurosci.org)
Each subunit has a large intracellular loop between the third and fourth transmembrane domains that is the likely site for regulated interactions with postsynaptic scaffolding proteins. (jneurosci.org)

A single set of standards contains markers for both visual gel and blot alignment plus TRF detection. (moleculardevices.com)
The ScanLater ™ Western Blot Protein Ladder is an essential component of the ScanLater ™ Western Blot Detection System. (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 blot is read with the ScanLater ™ Western Blot Detection Cartridge on the SpectraMax ® i3 or SpectraMax ® Paradigm ® Multi-Mode Microplate Readers. (moleculardevices.com)
They are detected by incubating the blot with Europium-labeled streptavidin and they show as light colored bands when read with the ScanLater Western Blot Detection Cartridge (Figure 1, left). (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)
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)
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)
Both the target protein and the normalization control need to fluoresce within the dynamic range of detection. (wikipedia.org)
When fluorescent detection is not utilized, the loading control protein and the protein of interest must differ considerably in molecular weight so they are adequately separated by gel electrophoresis for accurate analysis. (wikipedia.org)
Membranes need to be stripped and re-probed using a new set of detection antibodies when detecting multiple protein targets on the same blot. (wikipedia.org)
Because TPN is not dependent on a single loading control, validation of controls and stripping/reprobing of blots for detection of HKPs is not necessary. (wikipedia.org)
Replacements L38E and L41E, involving charge acquisition at residues predicted to contribute to the hydrophobic interface, reduced the dimerization signal in the protein ligation assay and prevented the detection of dimer/multimer species in both transiently expressed 3A proteins and in synthetic peptides reproducing the N terminus of 3A. (asm.org)

Methods We assessed the functional status of platelets by performing ELISA, western blot and RNA sequencing in a cohort of 71 patients and 68 age- and sex-matched healthy control subjects. (bmj.com)
The VEGF-A protein and mRNA levels in conditioned medium of RGC-5 cells incubated with AGE-modified BSA (AGE-BSA) were examined with real-time PCR and enzyme-linked immunosorbent assay (ELISA), and BSA-treated cells were used as controls. (molvis.org)
ABSTRACT 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)
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)

Moreover, the inhibition of DNA damage repair proteins Ku86 and B23 after proadifen treatment indicate a possible role of proadifen in DNA repair blockage, thus suppressing the reparation rate of MTX-induced DSBs. (spandidos-publications.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)
Dying cells have been defined as apoptotic by distinguishing features, including cell contraction, nuclear fragmentation, blebbing, apoptotic body formation and maintenance of intact cellular membranes to prevent massive protein release and consequent inflammation. (nature.com)

These results indicate that limited membrane permeabilization occurs in blebs and apoptotic bodies before secondary necrosis, leading to acute and localized release of immunomodulatory proteins during the early phase of active apoptotic membrane blebbing. (nature.com)
Normalization occurs directly on either the gel or the blotting membrane. (wikipedia.org)
When using protein stains, the membrane may be incubated with the chosen stain before or after immunodetection, depending on the type of stain. (wikipedia.org)
At the neuromuscular junction, the acetylcholine receptor (AChR) is specifically clustered in the postsynaptic membrane via interactions with rapsyn and other scaffolding proteins. (jneurosci.org)
The mechanism by which MuSK signaling regulates the interaction of the AChR with scaffolding proteins that localize it in the postsynaptic membrane, however, remain unknown. (jneurosci.org)

The pathology is caused by a mutation in the huntingtin gene which results in the production of an abnormal protein, mutant huntingtin (mHtt). (bmj.com)
Scope includes mutations and abnormal protein expression. (cancerindex.org)

The viral particle is composed of a protein capsid that contains a positive-sense RNA molecule of about 8,500 nucleotides that is infectious and encodes a single polyprotein, which is processed in infected cells by cis - and trans -acting viral proteases ( 55 ) to yield different polypeptide precursors and the mature viral proteins ( 9 , 62 ). (asm.org)
The viral genome encodes four structural capsid proteins (VP1 to VP4) and seven nonstructural (NS) proteins, the leader Lb/ab protease, and proteins encoded in the P2 (2B and 2C) and P3 (3A, 3B, 3C, and 3D) regions ( 9 ). (asm.org)

Western blot shows an increase in HSP70 that occurs when cells are exposed to temperatures that are higher than normal. (moleculardevices.com)

Lipids, polysaccharides and nucleic acids bind to proteins which increases the viscosity of the samples and often results in horizontal streaking on the 2D-PAGE. (hindawi.com)
However, it remains unclear where these proteins bind on the AChR and how the interactions are regulated. (jneurosci.org)

In this work, FMDV 3A homodimerization was evidenced by an in situ protein fluorescent ligation assay. (asm.org)

As we previously described, proadifen, a P450 monooxygenase inhibitor, might also be able to inhibit some ABC transporters, including breast cancer resistance protein (BCRP). (spandidos-publications.com)
More than 40 ABC transporters have been identified so far, but just a few play a role in MDR, including the breast cancer resistance protein (BCRP/ABCG2). (spandidos-publications.com)

Normalization of Western blot data is an analytical step that is performed to compare the relative abundance of a specific protein across the lanes of a blot or gel under diverse experimental treatments, or across tissues or developmental stages. (wikipedia.org)

What does this gene/protein do? (cancerindex.org)
What pathways are this gene/protein implicaed in? (cancerindex.org)
Histidine-rich protein 2/3 antigen concentrations were reduced with corresponding gene deletions. (cdc.gov)

Tissues are homogenized in protein lysis buffer supplemented with a cocktail of 7 mammalian protease inhibitors to minimize proteolysis, precisely quantified, and stored at -80oC. (delos.info)

The ScanLater Western Blot workflow follows standard gel loading and blotting methods up to the secondary antibody incubation step. (moleculardevices.com)
Eu-streptavidin binds to the protein ladder, and Eu-secondary antibody binds specifically to the primary antibody attached to the protein of interest. (moleculardevices.com)
Since HKP levels can be inconsistent between tissues, scientists can control for the protein of interest by spiking in a pure, exogenous protein of a known concentration within the linear range of the antibody. (wikipedia.org)
The integrity of protein is tested by immunoblotting using specific beta-actin antibody. (delos.info)
Highly sensitive and specific recombinant antibody microarrays allowing screening of crude protein extracts are also being developed [ 14 ]. (hindawi.com)

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)
The linear relationship between the molecular weight of the biotinylated ladder proteins and their migration distance on the gel is examined in Figure 2 in comparison with a reference standard. (moleculardevices.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)
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. (novusbio.com)

When using fluorescent antibodies to image proteins in western blots, normalization requires that the user define the upper and lower limits of quantitation and characterize the linear relationship between signal intensity and the sample mass volume for each antigen. (wikipedia.org)

However, recent studies have shown that expression of housekeeping proteins (HKPs) can change across different cell types and biological conditions. (wikipedia.org)
2D-PAGE for proteome studies gives a map of proteins which reflect changes in protein expression levels, as well different isoforms and posttranslational modifications. (hindawi.com)
The expression of tumour necrosis factor (TNF)-α, interleukin (IL)-6, zonula occludens (ZO)-1, Beclin-1 and autophagy-related (atg) 5 mRNA was evaluated by quantitative PCR, and the expression of ZO-1, microtubule-associated proteins 1A/1B light chain 3B (LC3B) I and, LC3B II protein by Western blot. (karger.com)
This protein was produced in an in vitro wheat germ expression system that should preserve correct conformational folding that is necessary for biological function. (novusbio.com)
Proadifen also decreased the expression of γH2AX, which is involved in the recruitment of reparation proteins. (spandidos-publications.com)
We examined not just the effect of proadifen and MTX on the expression of BCRP, but we were also interested in other molecular mechanisms involved in the possible antitumour activity of proadifen alone and in combination with MTX, such as the expression of anti-apoptotic proteins, proteins involved in the regulation of BCRP and proteins involved in the reparation of chemotherapeutic drug-induced DNA damage. (spandidos-publications.com)

Fluorescent stains and stain-free gels require special equipment to visualize the proteins on the gel/blot. (wikipedia.org)

Glycation, the result of a protein or lipid molecule bonding with sugar molecules, is a consequence of the aging process. (molvis.org)

Along with protein samples of interest, simply load one of the lanes on the gel with 4 µL of ScanLater Western Blot Protein Ladder. (moleculardevices.com)
The overall goal of normalization is to minimize effects arising from variations in experimental errors, such as inconsistent sample preparation, unequal sample loading across gel lanes, or uneven protein transfer, which can compromise the conclusions that can be obtained from Western blot data. (wikipedia.org)

Currently, there are two methods for normalizing Western blot data: (i) housekeeping protein normalization and (ii) total protein normalization. (wikipedia.org)

Whereas the G s /G i -protein-dependent tuning of striatal neurons is fairly well known, the precise impact and underlying mechanism of G q -protein-dependent signals remain poorly understood. (jneurosci.org)

We identified 231 proteins released from actomyosin contraction-dependent blebs and apoptotic bodies by adapted SILAC (stable isotope labeling with amino acids in cell culture) combined with mass spectrometry analysis. (nature.com)

To analyze this effect anatomically, we manipulated G q -protein-dependent signaling selectively in neurons belonging to the direct or indirect striatal pathway. (jneurosci.org)
Acute G q -protein activation in direct-pathway or indirect-pathway neurons produced an enhancement or a decrease, respectively, of activity-dependent parameters. (jneurosci.org)
In contrast, sustained G q -protein activation impaired the functionality of direct-pathway and indirect-pathway neurons and disrupted the behavioral performance and electroencephalography-related activity tasks controlled by either anatomical framework. (jneurosci.org)

Quality of protein as indicated by the absence of smear (no degradation) and sharpness and resolution of the protein bands is verified by denatured SDS-PAGE using Coomassie blue staining. (delos.info)

This protein is ubiquitously expressed and known to confer toxicity to multiple cell types. (bmj.com)
Western blotting is an important technique used in cell and molecular biology. (delos.info)
T-bet and STAT-4, two Th1 promoting transcription factors, and STAT-6 and GATA-3, transcription factors which govern T helper cell type 2 (Th2) cell polarisation, were examined in duodenal biopsies by western blotting. (bmj.com)
It was previously reported that proadifen (SKF-525A), a well-known cytochrome P450 monooxygenase inhibitor, not only has anti-proliferative potential in some cancer cell lines, but it is also able to inhibit BCRP and MRP1 transporter proteins ( 8 ). (spandidos-publications.com)

While it is possible that this protein could display some level of activity, the functionality of this protein has not been explicitly measured or validated. (novusbio.com)

Multidrug resistance caused by the overexpression of ABC transporter proteins in cancer cells remains a major obstacle limiting chemotherapy efficacy. (spandidos-publications.com)

First, the stained gel or blot is imaged, a rectangle is drawn around the target protein in each lane, and the signal intensity inside the rectangle is measured. (wikipedia.org)
The signal intensity obtained can then be normalized with respect to the signal intensity of the loading internal control detected on the same gel or blot. (wikipedia.org)
Ineffective stripping could result in a weak signal from the target protein. (wikipedia.org)
It could be difficult to completely eliminate signal from highly-abundant proteins, so it is recommended that one detects lowly-expressed proteins first. (wikipedia.org)

3b) Western blot showing an increase in HSP70. (moleculardevices.com)
Compared with the BSA controls, the RGC-5 cells incubated with AGE-BSA showed a dose- and time-dependent increase in VEGF-A mRNA and VEGF-A protein secretion in the supernatant, with the highest levels achieved at 24 h. (molvis.org)

Due to the large number of proteins, no protein array platform has yet been designed that covers the entire human placental proteome. (hindawi.com)

Foot-and-mouth disease virus (FMDV) nonstructural protein 3A plays important roles in virus replication, virulence, and host range. (asm.org)

The extracted protein is provided in vials at a concentration of 5mg/ml and shipped on dry ice. (delos.info)

Housekeeping genes and proteins, including β-Actin, GAPDH, HPRT1, and RPLP1, are often used as internal controls in western blots because they are thought to be expressed constitutively, at the same levels, across experiments. (wikipedia.org)
Many HKPs are expressed at high levels and are preferred for use with highly-expressed target proteins. (wikipedia.org)
We also measured the levels of phosphorylated tuberous sclerosis complex 2 (TSC2) protein and Parkin protein level in SH-SY5Y cells. (cancerindex.org)

On the protein level, out of 5,150 proteins identified, 393 were significantly upregulated and 227 were significantly downregulated. (frontiersin.org)

The method does not involve an enzymatic reaction, and as a result removes the inherent variability for protein quantitation . (moleculardevices.com)

In order to gain the best possible protein separation on 2D-PAGE, an optimized sample preparation protocol for placental proteins was developed. (hindawi.com)

These blue bands are seen on the electrophoresis gel and on the blot after transfer. (moleculardevices.com)
White bands are biotinylated protein used to estimate molecular weights of blotted proteins. (moleculardevices.com)
Dark bands are pre-stained protein used to monitor progress of protein electrophoresis, blot orientation, and assess transfer efficiency. (moleculardevices.com)

In total protein normalization (TPN), the abundance of the target protein is normalized to the total amount of protein in each lane. (wikipedia.org)
This can improve precision (down to 0.1 µg of total protein per lane), cost-effectiveness, and data reliability. (wikipedia.org)

The most enriched proteins released were the nucleosomal histones, which have previously been identified as damage-associated molecular pattern proteins (DAMPs) that can initiate sterile inflammatory responses. (nature.com)

Anatomy28

Organisms18

Diseases2

Chemicals and Drugs101

Analytical, Diagnostic and Therapeutic Techniques and Equipment43

Phenomena and Processes64

Disciplines and Occupations2

Technology, Industry, Agriculture5

Information Science3

Health Care3

Apoptosis5

Kinase6

Recombinant protein3

Isoelectric1

Intracellular4

Detection14

ELISA4

Inhibition1

Membranes2

Membrane5

Abnormal protein2

Encodes2

Occurs1

Bind2

Assay1

Breast cancer2

Analytical1

Gene3

Lysis1

Antibody5

Molecular weight5

Antigen1

Expression6

Gels1

Molecule1

Lanes2

Methods1

Mechanism1

Analysis1

Indirect3

Coomassie1

Cell4

Functionality1

Remains1

Signal4

Increase2

Array1

Important1

Concentration1

Levels3

Level1

Result1

Preparation1

Bands3

Total2

Previously1