Western blotting was performed as described previously [43 (link)]. The primary antibodies used were: anti-pp53 (1:1,000, CST), anti-p53 (1:1000, CST), anti-p21 (1:500, Santa Cruz), anti-p16 (Santa Cruz, sc-468), anti-caspase 3 (1:1000, CST), anti-PARP [poly(ADP ribose) polymerase] 1 (1:3000, ECTOMICS), anti-HMGA2 (1:5000, ECTOMICS), anti-caspase 9 (1:1000, Bioworld), anti-Bax and anti-Bcl2 (1:1000, CST), anti-caspase 2 (1:1000, KeyGEN), anti-γH2A (1:2000, Millipore) and anti-β-actin (1:10 000, Sungene).
Anti caspase 9
Manufactured by Bioworld Technology
Sourced in China
Anti-caspase 9 is a lab equipment product that is used to detect the presence and activity of caspase-9, a key enzyme involved in the apoptosis (programmed cell death) pathway. It provides a tool for researchers to study the role of caspase-9 in various biological processes and disease models.
Automatically generated - may contain errors
Lab products found in correlation
Ripa lysis buffer, by Beyotime (2 mentions)
Anti γh2a, by Merck Group (1 mentions)
Anti p16, by Santa Cruz Biotechnology (1 mentions)
Anti p21, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 3, by Proteintech (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Sds page, by Beyotime (1 mentions)
Anti gapdh, by Bioworld Technology (1 mentions)
Anti bcl 6, by Abcam (1 mentions)
Anti p53, by Proteintech (1 mentions)
Anti cyt c, by Bioworld Technology (1 mentions)
Anti p erk, by Bioworld Technology (1 mentions)
Rabbit anti caspase 3, by Cell Signaling Technology (1 mentions)
Bca protein quantification kit, by Beyotime (1 mentions)
Anti ras, by BD (1 mentions)
Anti erk, by Bioworld Technology (1 mentions)
Supersignal west femto maximum sensitivity substrate ecl, by Thermo Fisher Scientific (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
Anti nf κb, by Bioworld Technology (1 mentions)
Bca protein assay, by Beyotime (1 mentions)
4 protocols using anti caspase 9
1
Comprehensive Western Blot Analysis
2
Protein Expression Analysis via Western Blot
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RIPA lysis buffer was used to isolate proteins (Beyotime, Shanghai, China), and the Bradford method determine protein concentrations. Proteins (20 μg) were divided by SDS-polyacrylamide gel electrophoresis (SDS-PAGE, Beyotime) and transferred onto PVDF membranes (Millipore, USA). After 2h blocking in 5% bovine serum albumin (BSA, Vicmed, Xuzhou, China), the membranes were overnight incubated in primary antibody, diluted at 4°C in 5% BSA.
The primary antibodies were:anti-Bcl-6 (1:1000, Abcam, USA), anti-caspase3 (1:1000, Proteintech, USA), and anti-GAPDH (1:5000, Bioworld), anti-p53 (1:1000, Proteintech), anti-caspase9 (1:1000, Bioworld, Shanghai, China). Washing, then incubating the membranes with Horseradish Peroxidase (HRP) -conjugated rat anti- rabbit immunoglobulin (1/10000) (1:5000, Bioworld) for 1 h, diluted in 5% BSA. After washing, the secondary antibody was visualized by an enhanced chemiluminescence kit (Vicmed).
The primary antibodies were:anti-Bcl-6 (1:1000, Abcam, USA), anti-caspase3 (1:1000, Proteintech, USA), and anti-GAPDH (1:5000, Bioworld), anti-p53 (1:1000, Proteintech), anti-caspase9 (1:1000, Bioworld, Shanghai, China). Washing, then incubating the membranes with Horseradish Peroxidase (HRP) -conjugated rat anti- rabbit immunoglobulin (1/10000) (1:5000, Bioworld) for 1 h, diluted in 5% BSA. After washing, the secondary antibody was visualized by an enhanced chemiluminescence kit (Vicmed).
3
Protein Extraction and Western Blot Analysis in MCF-7 Cells
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MCF-7 cells were harvested by the use of ice-cold RIPA lysis buffer (Beyotime Biotechnology, Shanghai, China) and protein concentration was determined using the BCA protein quantification kit (Beyotime Biotechnology, Shanghai, China); western blotting was performed as previously described [19 (link)]. Primary antibodies of anti-Ac-H3, anti-SOD1, anti-CAT, anti-ERK, anti-p-ERK, anti-Cyt-C, and anti-caspase 9 were purchased from Bioworld Technology (Nanjing, China), anti-PARP, rabbit anti-caspase 3, and anti-Ac-H3 were purchased from Cell Signaling Technology, Danvers, USA, anti-RAS was purchased from BD, USA, and anti-GAPDH was purchased from Earthox, Millbrae, USA. The secondary antibodies were purchased from Biogot Biotechnology (Nanjing, China), and ECL SuperSignal West Femto Maximum Sensitivity Substrate was purchased from Thermo Fisher.
4
Western Blot Analysis of Apoptotic Proteins
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Cells were lysed on ice and centrifuged at 15,000 r/min at 4°C for 20 min. Supernatants were collected and protein concentrations were measured by bicinchoninic acid (BCA) protein assay (Beyotime, China). All of the samples were boiled at 100°C for 5 min. Equal amounts of protein were resolved on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels and then transferred to nitrocellulose (NC) membranes. Membranes were blocked with 5% skim milk for 2 h at room temperature and then incubated with primary antibodies (anti-Bcl-2, anti-Bax, anti-caspase-9, anti-cytochrome c, anti-NF-κB, anti-Mcl-1, and anti-caspase-3 (Bioworld Technology, China); anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and anti-P53 (Santa Cruz Biotechnology, USA)) at 4°C overnight. The next day, NC membranes were washed three times with Tris-buffered saline with Tween 20 (TBST) and incubated with corresponding horseradish peroxidase (HRP)-conjugated secondary antibodies (anti-rabbit IgG and anti-mouse IgG; Bioworld Technology) at room temperature for 2 h. NC membranes were then washed three times with TBST and signals detected with SuperSignal ECL (Pierce, USA).
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