2/21/2023 0 Comments Beta actin western blot![]() ![]() Human skeletal muscle fiber type specific protein content. Galpin AJ, Raue U, Jemiolo B, Trappe TA, Harber MP, Minchev K, Trappe S. Housekeeping proteins: a preliminary study illustrating some limitations as useful references in protein expression studies. Ferguson RE, Carroll HP, Harris A, Maher ER, Selby PJ, Banks RE. The light-induced reactions of tryptophan with halocompounds. Comparison of Stain-Free gels with traditional immunoblot loading control methodology. Colella AD, Chegenii N, Tea MN, Gibbins IL, Williams KA, Chataway TK. The use of total protein stains as loading controls: an alternative to high-abundance single-protein controls in semi-quantitative immunoblotting. Aldridge GM, Podrebarac DM, Greenough WT, Weiler IJ. ![]() The novel SF technology adds lower variation to the results compared with the existing methods for correcting for loading inaccuracy in Western blotting of human skeletal muscle in applied physiology. In contrast, the RPs may not be affected in studies that include muscle wasting and differences in muscle fiber type. In conclusion, β-actin, GAPDH, and α-tubulin may not be used for normalization in studies that include subjects with a large age difference. Protein level of β-actin and GAPDH was lower in older men compared with young men. The methodological variation was relatively low in all four methods (4–11%). A stronger linearity of SF and β-actin compared with GAPDH and α-tubulin was observed. Furthermore, the inter- and intraindividual variation in signals obtained from SF and RP was measured in relation to ageing, muscle atrophy, and different muscle fiber type composition, respectively. ![]() The linearity of signal and the methodological variation coefficient was obtained. ![]() This was done using skeletal muscle samples from men subjected to physiological conditions often investigated in applied physiology where the intervention has been suggested to impede normalization (ageing, muscle atrophy, and different muscle fiber type composition). Therefore, three widely used reference proteins, as well as TP loaded measured by Stain-Free technology (SF) as normalization tool were tested. However, the signal sensitivity and the influence of physiological conditions may question the normalization methods. Immunofluorescent analysis of Hela cells, using beta Actin Antibody.Reference proteins (RP) or the total protein (TP) loaded is used to correct for uneven loading and/or transfer in Western blotting. Immunohistochemical analysis of paraffin-embedded (1) Human heart (2) Mouse testis (3) Human bladder cancer (4) Mouse heart, using beta Actin Antibody. 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. Specificity: beta Actin Antibody detects endogenous levels of total beta-Actin protein. Storage Condition and Buffer: 1 mg/ml in PBS, pH 7.4, containing 0.02% sodium azide and 50% glycerol. 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. However it should be noted that levels of beta-Actin may not be stable in certain cells. Therefore, Antibodies against beta-Actin are useful as loading controls for Western Blotting. 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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