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microbiology lab days 5 and 6 part 3 (biochemical assay, fermentation, beta galactosidase, onpg, TSI - microbiology lab days 5 and 6 part 4 (biochemical assay, fermentation, beta galactosidase, onpg, TSI
Quantitation of β-galactosidase activity. In yeast cells, co-transformed with pGADT7 (AD) and pGBKT7 (BD) constructs as indicated, β-galactosidase activity wa
Colorimetric and chemiluminescent assays for beta galactosidase, LacZ vectors, and a complete beta-galactosidase reporter system. Fast, simple assay for beta-galactosidase activity.
Colorimetric and chemiluminescent assays for beta galactosidase, LacZ vectors, and a complete beta-galactosidase reporter system. Fast, simple assay for beta-galactosidase activity.
Buy our Recombinant |em|E. coli |/em| beta Galactosidase protein. Ab79449 is a full length protein produced in Escherichia coli. Abcam provides free protocols…
This unit describes fixation and staining for b‐galactosidase activity; it has been successfully used on vertebrate embryos and tissue explants
Expression In Bacteria Of Beta-Galactosidase Fusion Proteins Carrying Antigenic Determinants Of The 2 X-Gene Products Of Bovine Leukemia- ...
|strong|Rabbit anti beta-galactosidase polyclonal antibody|/strong| detects beta-galactosidase, an inducible enzyme that catalyzes the hydrolysis of lactose and other beta-galactosides into monosaccha…
BACKGROUND: The enzyme beta-galactosidase present in the Kupffer cells of the liver has potential as a marker of liver dysfunction prior to transplantation. Spectrophotometric methods have insufficient sensitivity. METHODS: Fluorimetric methods have the required sensitivity and we have optimised such a method in a microtitre plate format to improve its utility. beta-galactosidase acts on the substrate 4-methylumbelliferyl-galactoside (MUG) to produce 4-methylumbelliferone (4-MU), detected fluorimetrically with excitation wavelength 355 nm and emission wavelength 460 nm. RESULTS: Reaction conditions in a citrate-phosphate buffer were optimised to give maximal enzyme activity: pH was optimal at 4.4 (range investigated 3.6-5.0) and substrate concentration at 3.33 mmol/l. A small specimen volume (10 microl) in 80 microl of substrate solution produced adequate fluorescent yield after an incubation period of 30 to 60 min at 37 degrees C. Reaction was terminated by addition of 200 microl of glycine-NaOH, pH 12
Beta-galactosidase molecule. Computer model showing the 4-chain-structure of bacterial beta-galactosidase. - Stock Image C035/6227
Anti Beta-Galactosidase (E. Coli) IgG-Biotin conjugate Antibody conjugate BGAL12-BTN Anti Beta-Galactosidase (E. Coli) IgG-Biotin conjugate Antibody conjugate BGAL12-BTN
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
1DP0: High resolution refinement of beta-galactosidase in a new crystal form reveals multiple metal-binding sites and provides a structural basis for alpha-complementation.
Alpha-complementation of β-galactosidase does not seem to yield activities equal to wildtype β-galactosidase. Depending on the fragment, the activity can be up to 24% of wildtype [5]. (If anyone has a better reference comparing results from a Miller assay of alpha-complementated β-galactosidase with wildtype, please include it here.) ...
METHODS AND RESULTS The method involves the harvest of autologous venous-derived endothelial cells, the efficient genetic modification of the cells through the use of recombinant retroviruses, and the subsequent implantation of the genetically modified cells on the surface of balloon-denuded arterial segments. With a rabbit model, freshly isolated endothelial cells were transduced with a recombinant retrovirus encoding the bacterial enzyme beta-galactosidase. The autologous transduced cells were then implanted on the surface of balloon-denuded ileofemoral arterial segments at different cell densities; after 1 to 14 days, the animals were killed, and the vessel segments were examined. Cells expressing the bacterial gene product, as determined by in situ staining for beta-galactosidase, were found to be present on the surface of 28 of the 32 arteries seeded with genetically modified cells. Vessels examined at 4 to 7 days after seeding displayed 40% to 90% coverage with transduced cells, even when ...
Beta-galactosidase, 0.1 ml. This gene encodes beta-galactosidase-1, a lysosomal enzyme that hydrolyzes the termil beta-galactose from ganglioside substrates and other glycoconjugates.
Beta-galactosidase, molecular model. This enzyme breaks down sugars containing galactose, such as lactose, into their basic units (monosaccharides). - Stock Image C015/1970
1DP0: High resolution refinement of beta-galactosidase in a new crystal form reveals multiple metal-binding sites and provides a structural basis for alpha-complementation.
The cae8 promoter is sensitive to the amino acid L-asparagine.The figure shows measured β-galactosidase activities from wild-type BW25113 cells, carrying the c
Several types of assays can be performed measuring galactosidase activity in yeast using 5-Bromo-4-chloro-3-indoxyl-α-D-fucopyranoside as subtrate.
p>The checksum is a form of redundancy check that is calculated from the sequence. It is useful for tracking sequence updates.,/p> ,p>It should be noted that while, in theory, two different sequences could have the same checksum value, the likelihood that this would happen is extremely low.,/p> ,p>However UniProtKB may contain entries with identical sequences in case of multiple genes (paralogs).,/p> ,p>The checksum is computed as the sequence 64-bit Cyclic Redundancy Check value (CRC64) using the generator polynomial: x,sup>64,/sup> + x,sup>4,/sup> + x,sup>3,/sup> + x + 1. The algorithm is described in the ISO 3309 standard. ,/p> ,p class="publication">Press W.H., Flannery B.P., Teukolsky S.A. and Vetterling W.T.,br /> ,strong>Cyclic redundancy and other checksums,/strong>,br /> ,a href="http://www.nrbook.com/b/bookcpdf.php">Numerical recipes in C 2nd ed., pp896-902, Cambridge University Press (1993),/a>),/p> Checksum:i ...
Beta-Galactosidase (E. Coli) Protein for Western Blot Western control BGAL11-C Beta-Galactosidase (E. Coli) Protein for Western Blot Western control BGAL11-C
C-terminal BetaGal reporter tag yeast expression plasmid. This vector allows the creation of reporter fusion proteins with no protease cleavage tag.
lacZ. No transcription. Structural genes lacZ. lacY. lac A. No transcription. Presence of lactose. Transcription and translation. Active regulator protein. Inactive regulator protein (repressor). Allolactose. 12 When lactose is present, some of it is converted into allolactose,.... RNA polymerase lacO operator. Transcription and translation. Active regulator protein. Inactive regulator protein (repressor) ...
Misc.Comments : The SK designation indicates the polylinker is oriented such that beta-galactosidase (lacZ) transcription proceeds through the SacI site first and the KpnI site last. pBluescript SK(+) carries an F1 origin of replication, oriented such that transcription proceeds in the same direction as beta-galactosidase (lacZ) transcription ...
p>The checksum is a form of redundancy check that is calculated from the sequence. It is useful for tracking sequence updates.,/p> ,p>It should be noted that while, in theory, two different sequences could have the same checksum value, the likelihood that this would happen is extremely low.,/p> ,p>However UniProtKB may contain entries with identical sequences in case of multiple genes (paralogs).,/p> ,p>The checksum is computed as the sequence 64-bit Cyclic Redundancy Check value (CRC64) using the generator polynomial: x,sup>64,/sup> + x,sup>4,/sup> + x,sup>3,/sup> + x + 1. The algorithm is described in the ISO 3309 standard. ,/p> ,p class="publication">Press W.H., Flannery B.P., Teukolsky S.A. and Vetterling W.T.,br /> ,strong>Cyclic redundancy and other checksums,/strong>,br /> ,a href="http://www.nrbook.com/b/bookcpdf.php">Numerical recipes in C 2nd ed., pp896-902, Cambridge University Press (1993),/a>),/p> Checksum:i ...
ATCC ® 87590™ Designation: YEplac112 TypeStrain=False Application: YE-type (episomal) shuttle vector vector permitting visual detection of recombinants beta-galactosidase beta-D-galactosidase
BioAssay record AID 404864 submitted by ChEMBL: Activation of human estrogen receptor beta expressed in HEK293 cells at 1 nM beta-galactosidase reporter gene assay relative to control.
Beta-galactosidases catalyze the hydrolysis of beta(1-3) and beta(1-4) galactosyl bonds in oligosaccharides as well as the inverse reaction of enzymatic condensation and transglycosylation. Here we report the crystallographic structures of Penicillium sp. beta-galactosidase and its complex with galactose solved by the SIRAS quick cryo-soaking technique at 1.90 A and 2.10 A resolution, respectively. The amino acid sequence of this 120 kDa protein was first assigned putatively on the basis of inspection of the experimental electron density maps and then determined by nucleotide sequence analysis. Primary structure alignments reveal that Penicillium sp. beta-galactosidase belongs to family 35 of glycosyl hydrolases (GHF-35). This model is the first 3D structure for a member of GHF-35. Five distinct domains which comprise the structure are assembled in a way previously unobserved for beta-galactosidases. Superposition of this complex with other beta-galactosidase complexes from several hydrolase ...
Substâncias complexas são convertidas, pela ação de enzimas, em moléculas solúveis, durante o processo de germinação, as quais são translocadas para a plântula em crescimento, servindo como fonte de energia ou estrutura física. Com o objetivo de quantificar a atividade enzimática da -galactosidase e a mobilização de mono e oligossacarídeos durante o período de germinação, foi conduzido este estudo utilizando-se sementes de jacarandá-da-bahia. As sementes foram mantidas em germinador a 25oC sob luz contínua, sendo avaliada a protrusão da radícula, pelo período de 10 dias. Foram quantificados os teores de mono e de oligossacarídeos, assim como a atividade específica da enzima -galactosidase nos tempos zero, um, três e cinco dias. Houve mobilização das reservas de glicose e manose nos cotilédones e xilose neste e no embrião nos três primeiros dias de germinação. A ramnose teve os teores aumentados nos cotilédones e no eixo embrionário. A rafinose foi o ...
Recombinant His-tagged mouse Fab fragment raised against beta-galactosidase. Original antibody is raised against beta-galactosidase. (RAB00031) - Products - Abnova
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You might also use the Kyoto Encyclopedia of Genes and Genomes, which has a lot of information conveniently summarized for many proteins; for example, the entry for the E. coli beta-galactosidase gene is here. Note that the entries for orthologies, pathways, motifs, cross-references to other databases, structures, etc. are all clickable.. ...
It was found that Concanavalin A (Con A) accelerates the rates of hydrolysis of E. coli beta-galactosidase and yeast invertase by binding to the product (glucose) formed in the reaction. The effect of Con A can be made ...
BioAssay record AID 142597 submitted by ChEMBL: Agonistic efficacy in human m1 muscarinic receptor which was expressed with the marker gene beta-galactosidase in NIH 3T3 cells. Maximal effect normalized to the effect of carbachol (%CCH)..
The presence of chromogenic substrate gives blue coloured colonies if the plasmid in the bacteria does not have an insert presence of insert results into insertional inactivation of Beta galactosidase and recombinant colonies do not produce any colour these are identified as Recombinant Colony
Cellular senescence is a fundamental cell fate playing significant and complex roles during tumorigenesis and natural aging process. However, the molecular determinants distinguishing senescence from other temporary and permanent cell-cycle arrest states such as quiescence and post-mitotic state and the specified mechanisms underlying cell-fate decisions towards senescence versus cell death in response to cellular stress stimuli remain less understood. In our studies, we aimed to employ multi-omics approaches to deepen our understanding of cellular senescence, in particular, regarding the specific molecular determinants distinguishing cellular senescence from other non-dividing cell fates. Notably, one of the most prominent features of cellular senescence differing from other non-dividing cell fates is the increased expression of senescence-associated beta-galactosidase. Because 5-Dodecanoylaminofluorescein Di-β-D-Galactopyranoside (C12FDG) is known as the substrate catalyzed by ...
A Q: for transfection gurus. We are measuring promoter activity using luciferase gene as a main reporter, fused to the putative promoter fragments, and CMV/beta-galactosidase gene as a second, normalizer activity. Both are introduced by lipofection into an aortic cell line. Luciferase activity is measured using a Promega kit, and the b-gal is measured via a chemiluminescence based kit from Tropix (GalactoLight). Before measuring the latter activity we heat-inactivate the endogenous activity. What we get is that for a given construct the luciferase activity in a 3X transfection series is relatively constant ( within a factor of 1.5), but the respective set of b-gal activities is variable within a factor of 3, which ruins the experiment. Any ideas? Has anyone used GalactoLight with success? -- Alexander Kraev, PhD Biochemie III, ETHZ Zurich Phone 41-1-632-31-47 Fax 41-1-632-12-13 e-mail kraev at bc.biol.ethz.ch ...
Using a fungus two-hybrid system, we isolated a book human centrosomal protein, CPAP (centrosomal P4. utilized to display screen for protein that connect to 4.1R-135. The top domains (HD; residues 1 to 209) of 4.1R-135 (4.1R-HD) was fused towards the GAL4 DNA-binding domains (GAL4-DB) in vector pAS2-1 (Clontech). This create was used as bait to display a Z-VAD-FMK inhibition human being lymphocyte cDNA library fused to a GAL4 activation website (GAL4-AD) in the pACT2 vector (Clontech). The two types of plasmids were then cotransformed into Y190, and the transformants were selected on SD minimal medium as previously explained (40). Positive colonies were further tested for -galactosidase activity using a colony-lift assay and liquid assay as explained by the manufacturer (Clontech). To thin down the head website region of 4.1R (4.1R-HD) that binds to CPAP, constructs containing numerous portions of 4.1R-HD were fused to GAL4-DB of the pAS2-1 vector (Fig. ?(Fig.1A).1A). The C terminus of CPAP ...
Dimerization data (column 1) represent relative binding to GST-TBP(181C) as determined in Figure 1E and are an average of six sets of data. See Figure 5 for standard errors. β-galactosidase activity (columns 3-6) is relative to wild-type test TBP in column 3 (1.0 = 3 Miller units using the high-sensitivity substrate CPRG) and represents an average of at least three independent determinations. All data were obtained in the linear range of the assay. The null allele expresses only the first 81 amino acids of TBP. "Test" HA-tagged TBP levels were determined by quantitative Western blotting with TBP antibodies as described in Figure 5B. TBP levels are relative to the endogenous (untagged) TBP (1 = 17,000 ± 2,000 molecules per cell). The standard error for all data is presented graphically in Figure 5E. Expression of the TBPEBmutants did not cause a decrease in the expression of TAFII145 (data not shown). "Low" in columns 3 and 7 indicates that the test TBP was driven by the TBP promoter; "high" in ...
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The results of this study are consistent with the following conclusions. First, unconjugated β-galactosidase is rapidly cleared from blood in vivo (Table 2), owing to rapid uptake of the unconjugated enzyme by liver and spleen (Figs. 2 and 3). Second, once inside cells, β-galactosidase is rapidly degraded in vivo such that 99% of the organ enzyme activity is lost at 4 h after an intravenous injection (Table 1). Third, the 116-kDa β-galactosidase (Fig. 1B) can be conjugated to the 8D3 TfRmAb without loss of enzyme activity (Fig. 1C). Fourth, there is minimal brain uptake of the unconjugated β-galactosidase, but there is a 10-fold increase in brain uptake of enzyme following conjugation to the 8D3 TfRmAb (Table 1; Figs. 2 and 3).. The β-galactosidase is rapidly removed from the blood due to the avid uptake of the enzyme by liver and spleen (Figs. 2 and 3), which confirms the earlier observation of Onodera et al. (1983). The blood concentration of the β-galactosidase-TfRmAb is 5- to 10-fold ...
ATCC ® 77267™ Designation: pUN65 TypeStrain=False Application: YC-type (centromeric) shuttle vector mutation detection shuttle vector vector permitting RNA synthesis in vitro vector permitting visual detection of recombinants beta-galactosidase beta-D-galactosidase
Edvotek 300 kit shows how to clone a DNA fragment by using ligation, transformation and an assay of ß-galactosidase. For advanced students.
Page contains details about β-galactosidase/ZIF-8 MOF coating . It has composition images, properties, Characterization methods, synthesis, applications and reference articles : nano.nature.com
Expresses the head domain of Nod fused to the coiled-coil domain of KHC and beta-galactosidase under the control of UAS; the fusion protein accumulates at the minus ends of microtubules, N.G ...
These are methods to screen for and assay the activity of the common reporter enzyme β-Glucuronidase (GUS) activity. Because the catalytic activity of β-Glucuronidase is very similar to β-Galactosidase (LacZ) these protocols are also very similar to the LacZ protocols. ...
reacts with native and denatured-reduced E. coli β-galactosidase(116 kD); may be usedfor detection of β-galactosidase expressed by E. colilacZ gene encoded in many cloned gene sequences,and serves as an indicator for fusion proteins encodedby an inserted DNA ...
In article ,leach-051093092638 at med-pharm5.bu.edu,, leach at mbcrr.harvard.edu (Martin Leach) writes: , In article ,andres.37.749754284 at calvin.jci.tju.edu,, , andres at calvin.jci.tju.edu (Andres Ferber) wrote: , ,, Does anybody know of a reference for staining mammalian cells transfected ,, with betagalactosidase constructs? ,, I am interested in staining the cells alive by adding Xgal to the culture ,, media without fixing the cells. Is it possible? Any suggestions or comments? ,, Andres Ferber , I have an old protocol that works well, instead of posting phone PROMEGA at , 1-800-356-9526 and they can fax you their protocol in their b-gal assay , booklet. , Martin Leach Hi, Ive done X-gal staining on fixed mamalian cells expressing lacZ, but can you really do it on live cells without fixing???? ^^^^^^^^^^ -- Cheers, Martin NNNN NN Martin A Kennedy (E-mail = mkennedy at chmeds.ac.nz) ZZZZZZZ NN NN NN Cytogenetic and Molecular Oncology Unit ZZZ NN NN NN Christchurch School of Medicine ZZZ ...