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XRCC1 is a DNA repair protein that plays a crucial role in the base excision repair (BER) pathway. It functions as a scaffold protein, coordinating the activities of multiple enzymes involved in the recognition, processing, and ligation of DNA lesions caused by oxidative stress, alkylation, or single-strand breaks. XRCC1 facilitates the recruitment and assembly of the BER machinery, contributing to the maintenance of genomic integrity.

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Lab products found in correlation

Parp 1, by Santa Cruz Biotechnology (2 mentions) X tremegene sirna transfection reagent, by Roche (2 mentions) Immobilon fl pvdf membrane, by Merck Group (1 mentions) Histone h2a, by Merck Group (1 mentions) Alexa fluor 680, by Thermo Fisher Scientific (1 mentions) α actin, by Abcam (1 mentions) Odyssey image analysis system, by LI COR (1 mentions) Histone h3, by Cell Signaling Technology (1 mentions) γh2ax, by Merck Group (1 mentions) Irdye 800, by Rockland Immunochemicals (1 mentions) γh2ax, by Cell Signaling Technology (1 mentions) Brca2, by R&D Systems (1 mentions) Cell lysis buffer, by Cell Signaling Technology (1 mentions) Ecl solution, by Thermo Fisher Scientific (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Protease inhibitor, by Roche (1 mentions) Rad51, by Abcam (1 mentions) Nucleofector system, by Lonza (1 mentions)

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Anti-XRCC1 (2 citations)

5 protocols using xrcc1

1

siRNA Knockdown of PARP-1, RAC1, XRCC1

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siRNA knockdown was performed using X-tremeGENE siRNA transfection reagent (Roche) and validated siRNA sequences (working concentration 200 nM) against PARP-1 (Santa Cruz), RAC1 (Millipore Sigma), and XRCC1 (Ambion) for 48 hours. A scramble sequence (Ambion) was used as a negative control.
Marcar L., Bardhan K., Gheorghiu L., Dinkelborg P., Pfäffle H., Liu Q., Wang M., Piotrowska Z., Sequist L.V., Borgmann K., Settleman J.E., Engelman J.A., Hata A.N, & Willers H. (2019). Acquired Resistance of EGFR-Mutated Lung Cancer to Tyrosine Kinase Inhibitor Treatment Promotes PARP Inhibitor Sensitivity. Cell reports, 27(12), 3422-3432.e4.
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2

siRNA Knockdown of PARP-1, RAC1, XRCC1

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siRNA knockdown was performed using X-tremeGENE siRNA transfection reagent (Roche) and validated siRNA sequences (working concentration 200 nM) against PARP-1 (Santa Cruz), RAC1 (Millipore Sigma), and XRCC1 (Ambion) for 48 hours. A scramble sequence (Ambion) was used as a negative control.
Marcar L., Bardhan K., Gheorghiu L., Dinkelborg P., Pfäffle H., Liu Q., Wang M., Piotrowska Z., Sequist L.V., Borgmann K., Settleman J.E., Engelman J.A., Hata A.N, & Willers H. (2019). Acquired Resistance of EGFR-Mutated Lung Cancer to Tyrosine Kinase Inhibitor Treatment Promotes PARP Inhibitor Sensitivity. Cell reports, 27(12), 3422-3432.e4.
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3

Western Blot Analysis of DNA Damage Response

Cited 1 time
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Whole cell extracts for western blotting were prepared as described previously (17 (link)). Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and transferred onto Immobilon-FL PVDF membranes (Millipore) according to standard procedures. Membranes were probed with the following antibodies: APE1 (Novus Biologicals, NB100–101), XRCC1 (Thermo Scientific, MS-1393-P0), α-actin (Abcam, ab6276), p21 (Cell Signaling Technology, 2947), p53 (Santa Cruz, sc-126), PAR (Trevigen, 4335-AMC-050), histone H2A.X phosphorylated at Ser139 (γH2AX, 05–636, Millipore), Sp1 (Millipore, 07–645), histone H3 (Cell Signaling Technology, 4499). Secondary antibodies conjugated with Alexa Fluor 680 (Molecular Probes) and IRDye® 800 (Rockland) fluorescent dyes were used. Detection and quantification was carried out using an Odyssey image analysis system (Li-Cor Biosciences).
Poletto M., Legrand A.J., Fletcher S.C, & Dianov G.L. (2016). p53 coordinates base excision repair to prevent genomic instability. Nucleic Acids Research, 44(7), 3165-3175.
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4

Cisplatin-Induced DNA Damage Response Signaling

Cited 3 times
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Control and IPF fibroblasts (n=8, each) used for cell viability assay (6 × 105 cells/60 mm cell culture dish) were treated with cisplatin for various time points. Cells were lysed with 1x cell lysis buffer (Cell Signaling technology, Beverly, MA) containing protease inhibitor (Roche Applied Science, Indianapolis, IN) and phosphatase inhibitor (Research Products International Corp., Mount Prospect, IL), and western blotting was performed as previously described (15 ,16 (link),17 (link)). The antibodies used for our study are: γH2AX (PRID: AB_2118009, Cell Signaling Technology) (21 (link)), H2AX (RRID: AB_10694556, Cell Signaling technology) (22 (link)), RAD51 (RRID: AB_2042762, Abcam, Cambridge, UK) (11 (link)), BRCA2 (RRID: AB_2259370; R&D systems, Minneapolis, MN) (11 (link)), pXRCC1 (phosphorylated at S518, T519, and T523; catalog No. ab195205; Abcam) (21 (link)), XRCC1 (RRID:AB_10985840; ThermoFisher Scientific, Pittsburgh, PA) (23 (link)), CK2α (catalog No. 2656; Cell Signaling technology)(24 (link)), PUMA (RRID:AB_10987708; Santa Cruz Biotechnology, Santa Cruz, CA)(25 (link)), or GAPDH (RRID:AB_627679; Santa Cruz Biotechnology) (26 (link)). The protein bands on a membrane were then detected by ECL solution (ThermoFisher Scientific) using Chemi Doc-IT2 image analyzer (UVP BioImage systems, Upland, CA), and quantified using VisionWorks LS program (UVP BioImage systems).
Im J, & Nho R.S. (2019). Fibroblasts from patients with Idiopathic Pulmonary Fibrosis are resistant to cisplatin-induced cell death via enhanced CK2-dependent XRCC1 activity. Apoptosis : an international journal on programmed cell death, 24(5-6), 499-510.
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5

Knockdown of BRCA1 and BRCA2 in MCF-7 Cells

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BRCA1 and BRCA2 small hairpin RNAs (shRNAs) were generated using a lentiviral pLKO.1-puromycin vector (Sigma Aldrich). The sequences used for the small hairpin RNA complementary to BRCA1 and BRCA2 were as follows:
For establishment of clonally-derived cell lines, MCF-7 cells were electroporated using nucleofector system (Amaxa) according to manufacturer’s directions. The transfected cells were seeded in 10-cm dishes containing complete DMEM media. After 72h, the plates were washed with PBS and media containing 3μg/ml puromycin was added. The individual puromycin-resistant cells were allowed to grow for another 4 weeks to form colonies. The colonies were picked by placing cloning cylinders around clearly separate colonies. Immunoblotting was used to select the BRCA1 and BRCA2 depleted clones. Clones were maintained in a media containing 3μg/ml puromycin. For transient transfection, we used 4μg of XRCC1, FEN1 siGenome SMART pool siRNA and Non-Targeting siGenome siRNA as a control (Thermo Scientific). Cells were electroporated using the nucleofector system (Amaxa), and were harvested 48h post electroporation.
Fridlich R., Annamalai D., Roy R., Bernheim G, & Powell S.N. (2015). BRCA1 and BRCA2 protect against oxidative DNA damage converted into double-strand breaks during DNA replication. DNA repair, 30, 11-20.
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