The following antibodies were used for western blotting: p53 (OP43; Merck Millipore), MDM2 (SMP14) (sc-965; Santa Cruz), AKT (2920; Cell Signaling Technology), phospho-AKT (Thr308) (9275; Cell Signaling Technology), phospho-AKT (Ser473) (9271; Cell Signaling Technology), PARP (9532; Cell Signaling Technology), caspase 3 (9661; Cell Signaling Technology), phospho-5′ AMP-activated protein kinase ([AMPK] bs-4002R; Bioss), AMPK (E-AB-30490; Elabscience Biotechnology Inc.), phospho-mammalian target of rapamycin ([mTOR] (E-AB-20929; Elabscience Biotechnology Inc.), mTOR (E-AB-32129; Elabscience Biotechnology Inc.), LC3A/B (4108; Cell Signaling Technology), α-tubulin (NB100-690; Novus Biologicals), GAPDH (10494-1-AP; Proteintech), HSP-40 (sc-59554; Santa Cruz Biotechnology), CHIP (sc-133066; Santa Cruz Biotechnology), and vinculin (GTX109749; GeneTex). For co-immunoprecipitation assay, cell lysates were incubated with p53 antibody (OP43, EMD Millipore, Billerica, MA) or HSP-40 antibody (sc-398766, Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C. Then samples were precipitated with appropriate protein A/G beads with matched IgG as negative control. The precipitants were analysed in SDS-PAGE western blot.
Gtx109749
Manufactured by GeneTex
GTX109749 is a laboratory equipment product. It is a piece of essential lab equipment used for various scientific applications. The core function of this product is to perform a specific task in the laboratory setting. Further details about its intended use or features are not available.
Automatically generated - may contain errors
Lab products found in correlation
P53 antibody, by Merck Group (1 mentions)
Hsp40, by Santa Cruz Biotechnology (1 mentions)
Sc 133066, by Santa Cruz Biotechnology (1 mentions)
α tubulin nb100 690, by Novus Biologicals (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
Phospho akt thr308, by Cell Signaling Technology (1 mentions)
Lc3a b, by Cell Signaling Technology (1 mentions)
Gapdh 10494 1 ap, by Proteintech (1 mentions)
Ab15277, by Abcam (1 mentions)
2 protocols using gtx109749
1
Western Blot and Co-Immunoprecipitation Assay
2
Capillary Gel Electrophoresis Western Immunoassay for Dystrophin Quantification
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Capillary gel electrophoresis Western immunoassay was performed using the Wes system (ProteinSimple) according to the manufacturer’s instructions using a 66–440 kDa Separation Module (ProteinSimple). Protein lysates were loaded into capillaries at a concentration of 0.2 mg/ml for detection and quantification of dystrophin and vinculin proteins. Here, we used anti-dystrophin (ab15277, Abcam, dilution 1:15) and anti-vinculin (GTX109749, GeneTex, dilution 1:100) antibodies together with anti-rabbit secondary antibodies (042-206, Protein Simple). A standard curve was generated using a mixture of WT/mdx samples denoting 20%, 10%, 7.5%, 5%, 2.5% and 0% WT dystrophin levels. Relative dystrophin expression was calculated based on dystrophin chemiluminescence intensities normalized to vinculin The area corresponding to chemiluminescence signal was determined using Compass for SW software. Dystrophin expression values were plotted along our standard curve to calculate % dystrophin (relative to WT) in mdx samples.
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