All the kits or reagents for western blotting were obtained from Elabscience (Wuhan, China). Cells were lysed using the RIPA lysis buffer, and the protein concentration was estimated using the BCA Protein Concentration Detection Kit (Catalog No. E-BC-K318). SDS-PAGE (10%; Catalog No. E-IR-R305) was employed to separate proteins, which were then transferred onto PVDF membranes. The membranes were blocked for 1.5 h in 5% skim milk. The membranes were then incubated with primary antibodies (anti-ANP: ab209232, 1:1,000; anti-BNP: ab236101, 1:2,000; anti-β-MHC: ab180779, 1:2,000; and anti-β-actin: ab8227, 1:5,000; Abcam, Cambridge, CA, USA) overnight at 4°C and then incubated with secondary antibodies (Catalog No. E-AB-1003; Elabscience) for 1 h at 25°C. Protein bands were visualized using an ECL luminous detection solution (Catalog No. E-BC-R347). Gray analysis was performed by normalizing to GAPDH levels.
Ab236101
Manufactured by Abcam
Sourced in United States
Ab236101 is a laboratory equipment product. It is designed for use in research and scientific applications. The core function of this product is to [description not available].
Automatically generated - may contain errors
Lab products found in correlation
Ab180779, by Abcam (1 mentions)
Ab209232, by Abcam (1 mentions)
Ab8227, by Abcam (1 mentions)
Ab81299, by Abcam (1 mentions)
Ab225844, by Abcam (1 mentions)
Ab150119, by Abcam (1 mentions)
Ab5245, by Abcam (1 mentions)
Pierce ecl system, by Thermo Fisher Scientific (1 mentions)
Nb100 2219, by Novus Biologicals (1 mentions)
Sc 9053, by Santa Cruz Biotechnology (1 mentions)
Hrp conjugated secondary antibody, by Bio-Rad (1 mentions)
Nb100 479, by Novus Biologicals (1 mentions)
3 protocols using ab236101
1
Western Blotting of Cardiac Markers
2
Comprehensive Cardiovascular Biomarker Analysis
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HXP (Batch: 15050101) was provided by Guangzhou Youcare Biopharmaceutics Co., Ltd. (Guangzhou, China). ANP (ab225844), BNP (ab236101), gamma H2A.X (γ‐H2A.X; ab81299), Cardiac Troponin T (ab8295), Anti‐DNA/RNA Damage antibody (ab62623), Goat polyclonal Secondary Antibody to Rabbit IgG ‐ H&L (Alexa Fluor® 488), pre‐adsorbed (ab150081), Goat polyclonal Secondary Antibody to Mouse IgG ‐ H&L (Alexa Fluor® 647), pre‐adsorbed (ab150119) was obtained from Abcam. Collagen‐I (14695–1‐AP), Collagen‐III (22734–1‐AP), ACTA‐1 (17521–1‐AP), ACTN2 (14221–1‐AP), GAPDH (60004–1‐Ig), TBP (22006–1‐AP) were purchased from Proteintech. β‐catenin (8480S), Phospho‐β‐Catenin (p‐β‐catenin; 9561S) were provided by CST. Goat anti‐Rabbit IgG Secondary Antibody (8715) and Goat anti‐Mouse IgG Secondary Antibody (6229) were purchased from SAB.
3
Cardiac Protein Detection and Quantification
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For GATA4, pGATA4 and BNP thirty μg and for MEF2C hundred μg of total protein from heart tissue was lysed in EBC + urea buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.5% NP-40, 3 M urea) resolved by SDS-PAGE, transferred to PVDF membrane, blotted, and probed with the following primary antibodies: anti-GATA4 pSer105 (1:600 ab5245, Abcam), anti-GATA4 (1:1000 sc-9053, Santa Cruz Biotechnology), anti-BNP (1:500 ab236101, Abcam), anti-vinculin (1:200 V4505, Merck) and anti-MEF2C (1:7500 sc-313×, Santa Cruz Biotechnology) followed by a HRP-conjugated secondary antibody (1:5000; Bio-Rad). For blotting of HIF1α and HIF-P4H-2 hundred ug of total protein from heart tissue lysed using EBC + urea lysis buffer and was resolved by SDS-PAGE, transferred with nitrocellulose membrane, blotted and probed with anti-HIF1α (1:500, NB100-479 Novus Biologicals) and anti-HIF-P4H-2 (1:500, Novus Biologicals NB100-2219)primary antibodies followed by HRP-conjugated secondary antibody. The Pierce ECL system (Ther-moScientific) was used for detection. Western blot images were quantified with fiji (ImageJ) software. For original blots see (Supplementary Fig. S1-S8).
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