Immunoprecipitation and western blot analysis were performed using the following antibodies: HA (Covance, MMS-101R), ERCC1 (Santa Cruz Biotechnology, FL297), XPF (Abcam, ab73720), anti-SLX4 (affinity purified rabbit serum immunized with SLX4 1-758), swine anti-rabbit (DEKO, P0399), and rabbit anti-mouse (DEKO, P0260).
Ercc1
Manufactured by Santa Cruz Biotechnology
Sourced in United States
ERCC1 is a protein involved in DNA repair processes. It plays a critical role in the nucleotide excision repair (NER) pathway, which removes a variety of DNA lesions, including those caused by UV radiation and certain chemotherapeutic agents. ERCC1 forms a heterodimer with the XPF protein and functions as an endonuclease, cleaving the damaged DNA strand to facilitate its removal and subsequent repair.
Automatically generated - may contain errors
Lab products found in correlation
Mms 101r, by Santa Cruz Biotechnology (1 mentions)
Enhanced chemiluminescence detection reagent, by Cytiva (1 mentions)
Vimentin, by Abcam (1 mentions)
Anti abcg2, by Santa Cruz Biotechnology (1 mentions)
E cadherin, by Abcam (1 mentions)
Ube2c, by Abcam (1 mentions)
Cleaved capase 3, by Abcam (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Anti par antibody, by Bio-Techne (1 mentions)
Rad51, by Cell Signaling Technology (1 mentions)
Anti uhrf1, by Santa Cruz Biotechnology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
γh2ax, by Cell Signaling Technology (1 mentions)
Brca1, by Abcam (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
5 protocols using ercc1
1
Immunoprecipitation and Western Blotting
2
Western Blotting Technique for Protein Analysis
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Western blotting was performed as previously described 23 , 26 . Primary antibodies specific to hnRNPK, XIAP‐associated factor 1 (XAF1), ERCC excision repair 4, endonuclease catalytic subunit (ERCC4), ERCC1, G0/G1 switch 2 (G0S2) (1:200; Santa Cruz Biotechnology), cyclin A2, cyclin D1, cyclin E2, cleaved caspase‐7, and GAPDH (1:1000; CST, Danvers, Massachusetts, USA) were used. The blots were then incubated with goat anti‐rabbit or anti‐mouse secondary antibody (CST) and visualized using enhanced chemiluminescence.
3
Western Blot Analysis of Protein Expression
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For western blotting analysis, proteins of each group cells were denatured at 100°C boiled 5 min with SDS loading buffer. The proteins were transferred to PVDF transfer membrane. Membranes were incubated with the indicated antibodies overnight at 4°C followed by immunoblotting analysis. Proteins were detected using enhanced chemiluminescence detection reagents (Amersham). Tubulin was internal control. The primary antibodies used in this study were 1:1000 rabbit anti-ABCG2 and ERCC1 (Santa Cruz, Dallas, TX, USA), 1:1000 antibody of Tubulin, UBE2C, ZEB1, ZEB2, vimentin, E-cadherin, and cleaved capase-3 (Abcam, Cambridge, UK). The gray intensity analysis of western blotting images was carried out by ImageJ software.
4
Protein Expression Analysis by Western Blot
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Protein levels were assessed by western blot. Antibodies against HDAC, PARP, cleaved-PARP, H3, Acetyl-H3, and RAD51 were purchased from Cell Signaling Technology (CST). Anti-UHRF1, Ku-70, ERCC1, MSH2, MSH6, GAPDH and anti-α-tublin antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-BRCA1 and phosphorylated-BRCA1(ser988) was purchased from ABclonal Technology (Upper Heyford, UK). Anti-PAR antibody was purchased from Trevigen (4335-MC-100, MD, USA).
5
Western Blot Protein Analysis Protocol
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Cells were lysed in 2X sample buffer on ice for 10 min. Proteins were denatured at 95 °C for 5 min and stored at −80 °C. Western blotting was performed using a standard protocol, and antibodies against XPF (Thermo Fisher Scientific, cat#MA5-12060, dilution 1:1,000), FANCM (Merck, cat#MABC545, dilution 1:1,000), γH2AX (Cell Signaling, cat#9718 T, dilution 1:1,000), BRCA1 (Abcam, cat#ab16780, dilution 1:1,000), SLX4 (Bethyl Laboratories, cat#A302-269A, dilution 1:1,000), CSB (Bethyl Laboratories, cat#A301-345A, dilution 1:1,000), BLM (Bethyl Laboratories, cat#A300-110A, dilution 1:4,000), mCherry (GeneTex, cat#GTX128508, dilution 1:5,000), tubulin (Santa Cruz, cat#sc-134239, dilution 1:5,000), RNase H1 (GeneTex, cat#GTX117624, dilution 1:1,000), ERCC1 (SantaCruz, cat#sc-17809, dilution 1:1,000) and GAPDH (Cell Signaling, cat#2118, dilution 1:5,000) were used. Antibody information is provided in Supplementary Data 1 .
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