Human BRCA1 C-Terminus Antibody

BRCA1 in Human Breast Cancer Tissue. BRCA1 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Human BRCA1 Monoclonal Antibody (Catalog # MAB22101) at 5 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

BRCA1 in Human Breast. BRCA1 was detected in immersion fixed paraffin-embedded sections of human breast array using Human BRCA1 Monoclonal Antibody (Catalog # MAB22101) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human BRCA1 by Western Blot. Western blot shows lysates of U2OS human osteosarcoma cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Human BRCA1 Monoclonal Antibody (Catalog # MAB22101) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for BRCA1 at approximately 240 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of BRCA1 C-Terminus by Western Blot Microtubule-dependent nuclear deformations and DSBs mobility. A Immunoblot for BRCA1, CHK2 and phospho-CHK2 (P-CHK2) in p53-deleted (sg-P53) Brca1F/F Mouse Embryonic Fibroblasts (MEFs) 72 h after transduction with Hit&Run Cre and/or 6 h treatment with PARPi (0.5 μM). Actin is shown as loading control. b Examples of 10 min traces of mCherry-BP1-2 foci in Brca1F/F MEFs 72 h after Brca1 deletion, 6 h after PARPi addition in the absence or presence of taxol (1 h) or nocodazole (2 h). Scale bars: 10 μm. c MSD of mCherry-BP1-2 foci in the indicated MEFs as described in (b), with SD. Total foci analyzed: 1618 for DMSO, 1056 for taxol, and 1085 for nocodazole from 35, 30, 28 nuclei, respectively from n = 3 independent experiment. d Representative image of Brca1F/F MEFs without any treatment or 72 h after Cre-mediated deletion of Brca1, PARPi treatment for 6 h treatment and/or incubation with taxol for 1 h. Highlighted boxes indicate nuclear deformations as vertical invaginations (blue), light (green) and deep (yellow) lateral invaginations, and longitudinal invaginations (magenta). In black and gray boxes, the control nuclear edges and nuclear interior with no invaginations, respectively. Scale bars: 2 μm. Magnification ×4.25. (e) Quantification of nuclear deformations as in (d) from 40, 20 and 22 cells for each condition derived from one representative experiment. Statistical analysis by Kruskal–Wallis test for multiple comparisons. (*) P < 0.05 (p = 0.0381); P ≧ 0.05 are not significant. Source data are provided as a Source Data file. See also Supplementary Fig. 1. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40527886), licensed under a CC-BY license. Not internally tested by R&D Systems.