(PubMed link indicates BioLegend citation) For consistent results, please use TBS/T buffer for Western blotting that contains 0.15 M NaCl as indicated in the BioLegend recommended protocol. The binding of this antibody to its target is sensitive to salt concentration. The optimal dilution should be determined by titration for each individual assay of interest.Ģ5 µl and 100 µl of Direct-Blot™ HRP antibody can be used for approximately 5 and 20 Western blots, respectively, at the recommended concentration/dilution. For Western blotting, the suggested dilution is 1:1000-1:2000. Upon receipt, the antibody solution should be stored undiluted at -20☌, and protected from prolonged exposure to light.Įach lot of this antibody is quality control tested by Western blotting. Lot-specific (to obtain lot-specific concentration, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.) The antibody was purified by affinity chromatography and conjugated with HRP under optimal conditions. Discovery and prioritization of somatic mutations in diffuse large B-cell lymphoma (DLBCL) by whole-exome sequencing.This antibody is provided in 50% glycerol in aqueous buffered solutions with preservatives.Beta-actin variant is necessary for Enterovirus 71 replication.Total Protein Analysis as a Reliable Loading Control for Quantitative Fluorescent Western Blotting.Western blot analysis of beta-actin in various tissues and cell lines using Proteintech antibody 66009-1-Ig at a dilution of 1:20000 (Exposure time: 10 seconds).īeta-actin has been implicated in hand, foot, and mouth disease by proteomics and large diffuse B-cell lymphoma (3, 4). This is essential in some experimental systems where it may not be possible to always collect the exact same number of cells for each sample, such as when working with primary cells, tissue samples, or systems where you may be altering protein synthesis, cell size/number, or apoptosis (1).įigure 3. In Western blotting, beta-actin (42 kDa) is commonly chosen as a loading control due to its broad and consistent expression across all eukaryotic cell types and the fact that expression levels of this protein are not affected by most experimental treatments (Figure 3).Beta-Actin runs as a “loading control” to show that very similar amounts of protein were loaded in each lane. They can be used to normalize the levels of protein detected by confirming that protein loading is the same across the gel. Loading controls are essential for proper interpretation of Western blots. Why is beta-actin commonly used as a control in Western blots? Since the immunogen for 60008-1-Ig is the internal sequence, this antibody can recognize a wide range of species and can be broadly used in research. However, the N-terminal sequence is less conserved among species than the C-terminus.
#Beta actin western blot full#
Proteintech’s monoclonal beta-actin antibody ( 60008-1-Ig ) (Figure 1) recognizes all fragments, while 66009-1-Ig antibody (Figure 2) can only detect the full length of beta-actin. Beta-actin is highly conserved among species. Though beta-actin has a molecular weight of 42 kDa, degradation by apoptosis will produce 38-41 kDa fragments on a Western blot.
Four of these isoforms are expressed primarily in striated and smooth muscle cells, whereas beta-actin and gamma-actin isoforms are ubiquitously expressed. Beta-actin – a housekeeping protein Why is beta-actin commonly used as a control in Western blots? beta-actin and human diseases Related products ReferencesĪctin is a major component of the cytoskeleton, with six known isoforms in mammals.
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