Brca1 antibody

Manufactured by Santa Cruz Biotechnology

The BRCA1 antibody is a laboratory tool used for the detection and analysis of the BRCA1 protein. BRCA1 is a tumor suppressor protein that plays a crucial role in DNA repair. The antibody can be used in various immunological techniques, such as Western blotting, immunohistochemistry, and immunoprecipitation, to identify and study the BRCA1 protein in biological samples.

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Lab products found in correlation

Fluorescein isothiocyanate (fitc), by Jackson ImmunoResearch (1 mentions) Alexa fluor 594 fluorochrome, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Alpha tubulin, by Santa Cruz Biotechnology (1 mentions) Anti ha agarose, by Merck Group (1 mentions) Anti flag m2, by Merck Group (1 mentions)

Spelling variants of this product

BRCA1 (61 citations) Anti-BRCA1 antibody (6 citations) Mouse BRCA1 [D9] antibody (2 citations) BRCA1 primary antibody (1 citations)

3 protocols using brca1 antibody

Measuring BRCA1 DNA Repair Foci

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Cells were plated directly on a coverslip and allowed to adhere with overnight incubation. The following day, the cells were irradiated and fixed with 4% paraformaldehyde for 10 minutes at room temperature. Cells were then washed with PBS and permeabilized with 70% ethanol overnight at 4°C and for 20 minutes with 0.1% Igepal at room temperature. Cells were washed, blocked with 2% BSA for 1 hour and incubated overnight with 1:1,000 Aurora Kinase A antibody (Cell Signaling Technology). Centrosomes were visualized by 1 hour incubation with a 1:500 AlexaFluor 594 fluorochrome (Invitrogen). Cells were then incubated with 1:500 alpha tubulin (Santa Cruz Biotechnology) for 1 hour at room temperature. Mitotic spindles were visualized by 1 hour incubation with 1:600 FITC (Jackson ImmunoResearch), and DNA was stained with 1 μg/mL DAPI (RRID:AB_2893474; Sigma). Pictures were captured with a Leica microscope.
BRCA1 foci were visualized by incubating 1:500 BRCA1 antibody overnight (Santa Cruz Biotechnology) and 1:600 FITC (Jackson ImmunoResearch) for 45 minutes at room temperature. Foci for at least one hundred cells were manually counted per experiment using ImageJ software. Each foci experiment was repeated at least once.

Molkentine D.P., Molkentine J.M., Bridges K.A., Valdecanas D.R., Dhawan A., Bahri R., Hefner A.J., Kumar M., Yang L., Abdelhakiem M., Pifer P.M., Sandulache V., Sheth A., Beadle B.M., Thames H.D., Mason K.A., Pickering C.R., Meyn R.E, & Skinner H.D. (2022). p16 Represses DNA Damage Repair via a Novel Ubiquitin-Dependent Signaling Cascade. Cancer Research, 82(5), 916-928.

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NSCLC Biomarker Expression Analysis

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A standard protocol was used for the immunohistochemistry (IHC) of the samples that were detected as NSCLC by hematoxylin and eosin staining. Briefly, formalin fixed, paraffin embedding, paraffin-embedded specimens, dewaxing to water, antigen repair, serum blocking, primary antibody incubation (BRCA1 antibody, Santa Cruz, SC-624; STMN1 antibody, CST, #13655; MAPT antibody, CST,#4019; TUBB3 antibody, CST, #5666), secondary antibody incubation, coloration, counterstaining, dehydration, block.
Each tissue specimen was evaluated independentlyby two pathologists, and eight random fields were used to assess the expression levels of BRCA1, STMN1, MAPT and TUBB3, and also to calculate an average score. In addition, the two pathologists were blinded to the clinical status of the patients. For each patient specimen, these biomarkers were assessed by distribution and intensity. The staining distribution of target proteins were evaluated with the percentage of stained cells, which was scored as 0-100 (low expression: 0-25, high expression: 26-100).

Wang M., Li W., Xing X., Zhang D., Lei J, & Li G. (2017). BRCA1 and STMN1 as prognostic markers in NSCLCs who received cisplatin-based adjuvant chemotherapy. Oncotarget, 8(46), 80869-80877.

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Immunoprecipitation of BRCA1 and USP4

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Cells were collected and lysed on ice for 30 minutes in NETENG-400 buffer containing 400 mM NaCl, 20 mM Tris-HCl, pH 7.4, 0.5 mM EDTA, 1.5 mM MgCl₂, 0.1% NP-40, and 10% Glycerol, with protease inhibitors and phosphatase inhibitors (Sigma and Selleck). The samples were centrifuged at 12,000 rpm for 10 min, and the supernatants were diluted with NETENG-0 (containing all the same ingredients as NETENG-400 except for without NaCl) by adding 1.67 volume of the supernatant. For immunoprecipitation, the lysates were incubated with 2 μg BRCA1 antibody (Santa Cruz, sc-6954) or 1 μg USP4 antibody (Proteintech, 66822) or normal mouse IgG overnight at 4 °C with rotation, and protein-A/G agaroses (Roche) were then added, and incubated for an additional 2–4 h. For tagged protein immunoprecipitation, protein samples and anti-Flag-M2 or Anti-HA Agarose (Sigma-Aldrich) were incubated at 4 °C with rotation for 6 h. The immunocomplexes were then washed with NETENG-150 buffer (containing the same ingredients as NETENG-400 except for with 150 mM NaCl). Both the lysates and eluates were examined by WB with the indicated antibodies.

Guo X., Ma Y., Zhang T., Liu R., Chang F., Yan X., Yu T., Wu P., Li Q., Xu L., Duan J., Li L., Su Y, & Shao G. (2024). The deubiquitinating enzyme USP4 regulates BRCA1 stability and function. NPJ Breast Cancer, 10, 35.

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