Cells were washed with cold PBS and lysed in RIPA buffer. Protein concentration was tested with the BCA Assay Kit (Bio-Rad, Richmond). Samples were denatured at 95 °C for 5 min, separated using SDS-PAGE, and transferred to polyvinylidene difluoride (PVDF) membranes (GE Healthcare, Piscataway, NJ, USA). After blockage with 5% BSA for 1 h at room temperature, the membrane was probed with rabbit polyclonal anti-FoxC1 (Abcam, 1:1000) overnight at 4 °C, then incubated with HRP-conjugated secondary antibody for 1 h, was visualized using the chemiluminescence system (Millipore), and exposed to X-ray medical films (Kodak/Carestream Health). Actin was used as a loading control.
Anti foxc1
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti-FoxC1 is a primary antibody that specifically recognizes the FoxC1 protein. FoxC1 is a transcription factor involved in various developmental processes. This antibody can be used for applications such as Western blotting and immunohistochemistry to detect and analyze the expression of FoxC1 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions)
Bca assay kit, by Bio-Rad (1 mentions)
Pvdf membrane, by GE Healthcare (1 mentions)
Anti h3k27me3, by ABclonal (1 mentions)
Bca assay kit, by Thermo Fisher Scientific (1 mentions)
Anti histone h3, by Bioworld Technology (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti beclin 1, by Santa Cruz Biotechnology (1 mentions)
Anti elavl1, by Abcam (1 mentions)
Anti gpx4, by Abcam (1 mentions)
Anti p62, by Abcam (1 mentions)
Anti lc3, by Abcam (1 mentions)
3 protocols using anti foxc1
1
Western Blot Analysis of FoxC1 Protein
2
Protein Isolation and Western Blot Analysis
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Protein isolation from whole cerebral cortex or cells was performed as descried previously33 (link). The concentration of protein was assessed by BCA assay kit (Thermo Fisher). Western blots were performed using the standard SDS-polyacrylamide gel electrophoresis method and electrotransferred onto PVDF membranes (Millipore, Billerica, MA, USA). After blocking with 3% (w/v) BSA (SunShine, Nanjing, China) in PBS for 2 h, the membranes were incubated with primary antibodies at 4 °C overnight, and then incubated with the corresponding secondary antibodies (CST, Boston, MA, USA; 1:10,000) for 1 h at room temperature. At last, the immunoblot was visualized and quantified by a gel densitometric scanning and analysis system (Bio-Rad, Berkeley, CA, USA).
Primary antibodies were used as follows: anti-SIRT6 (#12486; CST; 1:1000), anti-H3K9Ac (#9469; CST; 1:1000), anti-H3K56Ac (#4232; CST; 1:1000), anti-EZH2 (#5246; CST; 1:1000), anti-FOXC1 (ab227977; abcam, Cambridge, UK; 1:1000), anti-Pan Ac-K (A2391; abclonal, Wuhan, Hubei, China; 1:1000), anti-H3K14Ac (A7254; abclonal; 1:1000), anti-H3K27me3 (A2363; abclonal; 1:1000), anti-Histone H3 (BS90642; Bioworld Technology, Louis Park, MN, USA; 1:1000), anti-β-Actin (23660-1-AP; Proteintech, Wuhan, Hubei, China; 1:10,000).
Primary antibodies were used as follows: anti-SIRT6 (#12486; CST; 1:1000), anti-H3K9Ac (#9469; CST; 1:1000), anti-H3K56Ac (#4232; CST; 1:1000), anti-EZH2 (#5246; CST; 1:1000), anti-FOXC1 (ab227977; abcam, Cambridge, UK; 1:1000), anti-Pan Ac-K (A2391; abclonal, Wuhan, Hubei, China; 1:1000), anti-H3K14Ac (A7254; abclonal; 1:1000), anti-H3K27me3 (A2363; abclonal; 1:1000), anti-Histone H3 (BS90642; Bioworld Technology, Louis Park, MN, USA; 1:1000), anti-β-Actin (23660-1-AP; Proteintech, Wuhan, Hubei, China; 1:10,000).
3
Myocardial Protein Expression Analysis
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RIPA lysis buffer (ThermoFisher, MI, USA) was utilized to extract proteins from left ventricular myocardium tissues or cells according to standard protocol. Protein concentration of each sample was measured by using Pierce™ BCA Protein Assay Kit (ThermoFisher, MI, USA). Equal amounts of protein were loaded into SDS–polyacrylamide gels and separated through electrophoresis. Later the proteins were transferred from the gels to PVDF membranes (Sigma-Aldrich, USA). The membranes were blocked with 3% BSA for half an hour at room temperature and then incubated with primary antibodies overnight at 4 °C. On the next day the membranes were washed with TBST 3 times before incubation with specific secondary antibodies for 1 h at room temperature. Signals were detected by using the standard ECL kit. Primary antibodies used in the study were: Anti-ELAVL1 (1:1000; Abcam, USA); Anti-GPx4 (1:5000; Abcam); Anti-ferritin heavy chain (FTH1, 1:1000; Abcam); Anti-Beclin-1 (1:1000; Santa Cruz, USA); Anti-p62 (1:1000; Abcam); Anti-LC3 (1:3000; Abcam); Anti-FOXC1 (1:1000; Abcam); Anti-β-actin (1: 1000; Santa Cruz).
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