Im doing a standard, semi dry Western Blot using beta actin as a marker, but every time I am seeing a double banding pattern. These findings demonstrate that β-actin is present in human plasma and may possibly be used as a suitable loading control for plasma-based Western blot analysis in major depressive disorder.Ĭopyright © 2012 Elsevier Inc. Beta-actin, double bands - Problems with my western blot. Moreover, the total coefficient of variation for β-actin expression in the two groups was 9.2☑.2%. The data revealed no statistically significant difference between these two groups. Furthermore, the expression of plasma β-actin in major depressive disorder subjects and healthy controls was compared. In addition, the linearity of β-actin immunostaining and loaded protein amount was evaluated by Western blot, and a fine linearity (R²=0.974☐.012) was observed. In this study, an enzyme-linked immunosorbent assay was used to determine the concentration of β-actin in human plasma, which is 6.29☐.54 ng/ml. However, β-actin's presence in human plasma and this protein's putative role as a plasma-based loading control for Western blot analysis remain unknown. On account of its relatively high and constant expression, β-actin has been widely employed in Western blot of cell cultures and tissue extracts. Please note this antibody is raised against the b-nonmuscle. For normalization of protein levels in Western blot, a suitable loading control is required. This antibody detects a single clean band at just above 40kD that corresponds to beta Actin. Immunofluorescent analysis of Hela cells, using beta Actin Antibody.Western blot analysis is a commonly used technique for determining specific protein levels in clinical samples. View specifications, prices, citations, reviews, and more. Immunohistochemical analysis of paraffin-embedded (1) Human heart (2) Mouse testis (3) Human bladder cancer (4) Mouse heart, using beta Actin Antibody. Compare beta actin Western Blot Antibody Products from leading suppliers on Biocompare. Western blot analysis of beta Actin expression in (1)HeLa cell lysate (2)Human fetal kidney lysate (3) NIH/3T3 cell lysate (4) PC-12 cell lysate. Immunogen: A synthesized peptide derived from human beta Actin. It is built on a technique that involves transferring, also known as blotting, proteins separated by electrophoresis from the gel to a membrane where they can be visualized specifically. Specificity: beta Actin Antibody detects endogenous levels of total beta-Actin protein. Western blot Western Blot (WB) is a common method to detect and analyze proteins. Storage Condition and Buffer: 1 mg/ml in PBS, pH 7.4, containing 0.02% sodium azide and 50% glycerol. AM1021b was diluted at 1:1000 at each lane. For example, expression of beta-Actin in adipose tissue is very low and therefore it should not be used as loading control for these tissues. Western blot analysis of lysates from HepG2, HL-60 cell line (from left to right), using Beta-actin Antibody(Cat. However it should be noted that levels of beta-Actin may not be stable in certain cells. Its molecular weight is approximately 42 kDa. Its transcript is also commonly used as a housekeeping gene standard in qPCR. Therefore, Antibodies against beta-Actin are useful as loading controls for Western Blotting. Beta actin is often used in Western blotting as a loading control, to normalize total protein amounts and check for eventual protein degradation in the samples. The actin molecules found in cells of various species and tissues tend to be very similar in their immunological and physical properties. Beta-Actin is one of six different actin isoforms that have been identified.
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