Myocardial tissues were lysed in radioimmunoprecipitation buffer (Beyotime, Shanghai, China). The protein supernatants were extracted from the homogenized tissues by centrifugation at 12,000 g for 1 h. Protein concentration was determined using bicinchoninic acid method (Pierce, Rockford, IL, USA). Protein samples were performed with electrophoresis and transferred into polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% fat-free milk and probed with primary antibodies against Bax (1:2,000), Bcl-2 (1:2,000), collagen I (1:2,500), collagen III (1:2,500), p38 (1:3,000), p-p38 (1:3,000), ERK1/2 (1:3,500), p-ERK1/2 (1:3,500) and GAPDH (1:4,000) (Bioss, Beijing, China). Following incubation with horseradish peroxidase-conjugated immunoglobulin G (1:5,000; Abcam, Cambridge, MA, USA), the blots were detected by enhanced chemiluminescence (KeyGen, Nanjin, China).
Horseradish peroxidase conjugated immunoglobulin g
Manufactured by Abcam
Sourced in United States, China
Horseradish peroxidase-conjugated immunoglobulin G is a laboratory reagent used in various immunoassay techniques. It consists of immunoglobulin G (IgG) molecules that have been conjugated to the enzyme horseradish peroxidase. This conjugate can be used to detect and quantify target analytes in samples.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence, by Keygen Biotech (2 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Gapdh, by Bioss Antibodies (1 mentions)
Bcl2 associated x (bax), by Bioss Antibodies (1 mentions)
Erk1 2, by Bioss Antibodies (1 mentions)
P erk1 2, by Bioss Antibodies (1 mentions)
Radioimmunoprecipitation buffer, by Beyotime (1 mentions)
Anti sox2, by Abcam (1 mentions)
Anti nanog, by Abcam (1 mentions)
Anti oct4, by Abcam (1 mentions)
Anti fak, by Abcam (1 mentions)
Anti p fak, by Abcam (1 mentions)
Anti ube2t, by Abcam (1 mentions)
Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Anti grp78, by Abcam (1 mentions)
Ripa buffer, by Dingguo (1 mentions)
2 protocols using horseradish peroxidase conjugated immunoglobulin g
1
Myocardial Protein Quantification and Immunoblotting
2
Protein Expression Analysis of Tumor Tissues
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RIPA buffer (Ding Guo Chang Sheng Biotech, Beijing, China) was used to lyse the tumor tissues and cells on ice. The protein concentration of the supernatants collected from the lysates was calculated using a bicinchoninic acid protein assay kit (Pierce Biotechnology, Rockford, IL, USA). Samples (30 µg) were separated by SDS-PAGE and then transferred to PVDF membrane. The membrane was probed with anti-UBE2T and anti-GAPDH (1:2,000; Abcam, Cambridge, MA, USA), anti-FAK and anti-p-FAK (1:2,500; Abcam), anti-SOX2 and anti-Oct4 (1:3,000; Abcam), anti-Nanog, or anti-GRP78 (1:4,000; Abcam). Following incubation with horseradish peroxidase-conjugated immunoglobulin G (1:5,000; Abcam), the blots were detected by enhanced chemiluminescence (KeyGen, Nanjin, China).
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