Nb100 177

Manufactured by Novus Biologicals

NB100-177 is a laboratory equipment product offered by Novus Biologicals. It is designed for general laboratory use, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach without extrapolation.

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Lab products found in correlation

Horseradish peroxidase conjugated anti mouse immunoglobulin g, by Cell Signaling Technology (2 mentions) Anti rabbit immunoglobulin g, by Cell Signaling Technology (2 mentions) Ab213, by Abcam (1 mentions) Ab6326, by Abcam (1 mentions) Bn 34, by Cell Signaling Technology (1 mentions)

3 protocols using nb100 177

Antibody Panel for SIRF, PLA, and DNA Fiber Assays

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Antibodies used in SIRF and PLA assays are as follows: mouse anti-biotin (Sigma-Aldrich, BN-34), rabbit anti-biotin (Cell Signaling Technology, D5A7), rabbit anti-XRCC3 (Abnova, PAB24835), mouse anti-RAD51C (Abnova, H00005889-M01), rabbit anti-RAD51C (Abcam, ab72063), and mouse anti-RAD51 (Abcam, ab213). Antibodies used in immunoblotting and immunoprecipitation are mouse anti-RAD51C (Novus, NB100-177) and rabbit anti-XRCC3 (Novus, NB100-165). Antibodies used in DNA fiber assays are anti-IDU (BrdU, Beckton Dickinson, 347580, 1:50) and CldU (BrdU, Abcam, ab6326, 1:100). More detailed antibody information can be found in the Supplementary Table 3. Uncropped and unprocessed scans of Western blots and gels can be found in the Source data file.

Longo M.A., Roy S., Chen Y., Tomaszowski K.H., Arvai A.S., Pepper J.T., Boisvert R.A., Kunnimalaiyaan S., Keshvani C., Schild D., Bacolla A., Williams G.J., Tainer J.A, & Schlacher K. (2023). RAD51C-XRCC3 structure and cancer patient mutations define DNA replication roles. Nature Communications, 14, 4445.

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Lysis and Immunoblotting of Cellular Proteins

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Cells were lysed in ice-cold cell lysis buffer (50 mM HEPES, 1% Triton X-100, 10 mM NaF, 30 mM Na₄P₂O₇, 150 mM NaCl, and 1 mM EDTA freshly supplemented with 10 mM β-glycerophosphate, 1 mM Na₃VO₄, 20 μg/ml pepstatin A, 10 μg/ml aprotinin, 20 μg/ml leupeptin, 40 μM microcystin-LR). Immunoblotting was done using the following primary antibodies: mouse monoclonal RAD51C (1:1000, catalog no. NB100-177, Novus Biologicals, RRID:AB_10001856), rabbit monoclonal XRCC1 (1:1000, catalog no. 3631-1, Epitomics, RRID:AB_10897570), rabbit monoclonal IRS1 (1:1000, catalog no. 3407T, Cell Signaling Technology, RRID:AB_2127860), rabbit polyclonal phospho-IRS1 (Tyr612) (1:1000, catalog no. 44-816G, Thermo Fisher Scientific, RRID:AB_2533768), rabbit monoclonal PARP1 (1:1000, catalog no. ab32138, Abcam, RRID:AB_777101), rabbit monoclonal histone H3 (1:1000, catalog no. 4499, Cell Signaling Technology, RRID:AB_10544537), and mouse monoclonal HSP90 (D. Toft, Mayo Clinic, H9010). Secondary antibodies used were horseradish peroxidase–conjugated anti-mouse immunoglobulin G (1:10,000, catalog no. 7076S, Cell Signaling Technology, RRID:AB_330924) and anti-rabbit immunoglobulin G (1:10,000, catalog no. 7074S, Cell Signaling Technology, RRID:AB_2099233).

Kanakkanthara A., Hou X., Ekstrom T.L., Zanfagnin V., Huehls A.M., Kelly R.L., Ding H., Larson M.C., Vasmatzis G., Oberg A.L., Kaufmann S.H., Mansfield A.S., Weroha S.J, & Karnitz L.M. (2021). Repurposing Ceritinib Induces DNA Damage and Enhances PARP Inhibitor Responses in High-Grade Serous Ovarian Carcinoma. Cancer Research, 82(2), 307-319.

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Cell Lysis and Immunoblotting for DNA Repair Proteins

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Cells were lysed in ice-cold cell lysis buffer (50 mmol/L HEPES, 1% Triton X-100, 10 mmol/L NaF, 30 mmol/L Na₄P₂O₇, 150 mmol/L NaCl, and 1 mmol/L EDTA freshly supplemented with 10 mmol/L β-glycerophosphate, 1 mmol/L Na₃VO₄, 20 μg/mL pepstatin A, 10 μg/mL aprotinin, 20 μg/mL leupeptin, 40 μmol/L microcystin-LR). Immunoblotting was done using the following primary antibodies: mouse monoclonal RAD51C (1:1,000, catalog no. NB100–177, Novus Biologicals, RRID:AB_10001856), rabbit monoclonal XRCC1 (1:1,000, catalog no. 3631-1, Epitomics, RRID:AB_10897570), rabbit monoclonal IRS1 (1:1,000, catalog no. 3407T, Cell Signaling Technology, RRID:AB_2127860), rabbit polyclonal phospho-IRS1 (Tyr612; 1:1,000, catalog no. 44-816G, Thermo Fisher Scientific, RRID:AB_2533768), rabbit monoclonal PARP1 (1:1,000, catalog no. ab32138, Abcam, RRID:AB_777101), rabbit monoclonal histone H3 (1:1,000, catalog no. 4499, Cell Signaling Technology, RRID:AB_10544537), and mouse monoclonal HSP90 (D. Toft, Mayo Clinic, H9010). Secondary antibodies used were horseradish peroxidase–conjugated anti-mouse immunoglobulin G (1:10,000, catalog no. 7076S, Cell Signaling Technology, RRID:AB_330924) and anti-rabbit immunoglobulin G (1:10,000, catalog no. 7074S, Cell Signaling Technology, RRID:AB_2099233).

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