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              技術文章

              BRCA1 antibody - ChIP grade Catalog Number:OM241912

              點擊次數:29 發布時間:2024/8/23 13:24:46
               

              Product Profile

              Product Name BRCA1 antibody - ChIP grade
              Antibody Type Primary Antibodies
              Product description
              Mouse monoclonal [6B4] to BRCA1BRCA1-6B4 recognizes full length BRCA1, a 220kDa nuclear phosphoprotein, and does not recognize the exon 11 splice variant. Mutations in this tumor suppressor gene greatly increase the risk of breast cancer. In a high proportion of breast and ovarian cancer cell lines, BRCA1 aberrantly mislocates to the cylasm. Recent studies suggest a role for BRCA1 in DNA double strand break repair because of its association with Rad51.
              More 
              Immunogen
              BRCA1 protein fragment expressed in E. coli corresponding to amino acids 341-748.

              Key Feature

              Clonality Monoclonal
              Isotype IgG1
              Clone Number 6B4
              Host Species Mouse
              Tested Applications

              ICC/IFIHCIPWBChIP

               
              Suggested starting dilutions are as follows: WB: 1:500-1:3000, ICC/IF: 1:100-1:1000, IHC, IP, ChIP. Not yet tested in other applications. Optimal working dilutions should be determined experimentally by the end user.:
              Species Reactivity

              HumanMouse

              Concentration 1 mg/ml
              Purification Protein G

              Target Information

              Gene Synonyms
              113705
              RNF53
              BRCA1
              BRCAI
              672
              PSCP
              BRCC1
              P38398
              IRIS
              BROVCA1
              PNCA4
              More 
              Target BRCA1
              Molecular Weight(MW) 220 kDa (note)
              Tissue Specificity This antibody recognizes BRCA1, a 220-kDa nuclear phosphoprotein, and does not recognize the exon 11 splice variant. Mutations in this tumor suppressor gene greatly increase the risk of breast cancer.
              Cellular Localization Nuclear
              Light Chain Type kappa

              Application

              • Application

                BRCA1 antibody detects BRCA1 protein by western blot analysis. Whole cell extracts (30 and 50 µg) was separated by 5% SDS-PAGE, and the membrane was blotted with BRCA1 antibody at a dilution of 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

              • Application

                BRCA1 antibody 6B4 was used for western blot assay at 1:1000 antibody dilution. western blot assay was performed using MCF7 (human) and MEF(mouse embryonic fibroblast) cell lysate separated in a 5-15% gradient gel. Observed molecular weight of BRCA1: 220 KD. Predicted molecular weight of BRCA1: 220 KD. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

              • Application

                 

              • Application

                BRCA1 antibody 6B4 was used for immunofluorescent staining of BRCA1 nuclear foci induced by ionizing radiation. IR-treated (2 hr /4 gray IR) U2OS cells were pre-extracted with CSK buffer on ice for 4 min before fixation with 4% PFA in room temperature, and then subjected to immunostaining. DAPI was used to counterstain nucleus. 6B4 was used at 1:400 dilution. Secondary antibody (Alexa Fluor-488) used for detection of primary antibody (BRCA1 antibody 6B4).

              • Application

                BRCA1 antibody 6B4 and BRCA1 antibody 17F8 was used for IP-WB assay. 6B4 alone (4 microgram), 17F8 alone (4 microgram), 6B4 plus 17F8 (2 microgram each), and mouse control normal IgG were used in an immunoprecipitation assay with MCF7 cell extract. Immunoprecipitated BRCA1 was detected in WB using BRCA1 antibody 6B4 at 1:1000 dilution. HeLa whole cell extract (20 microgram) was used as input in the Western blot assay.

              • Application

                BRCA1 antibody 6B4 and BRCA1 antibody 17F8 were used for ChIP assay. The 6B4 and 17F8 mixture (3 microgram each), or normal mouse IgG (6 microgram) were incubated with HeLa chromatin extract (100 microgram each) in the ChIP assay. Enrichment of genomic DNA on a BRCA1 target gene promoter (HMGA2) was validated by a Q-PCR assay.

              • Application

                Non-transfected (–) and transfected (+) 293T whole cell extracts (60 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with BRCA1 antibody [6B4] - ChIP grade diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

              Positive Control 293T , MCF7 , MEF
              Application Notes
              Suggested starting dilutions are as follows: WB: 1:500-1:3000, ICC/IF: 1:100-1:1000, IHC, IP, ChIP. Not yet tested in other applications. Optimal working dilutions should be determined experimentally by the end user.:

              Additional Information

              Form Liquid
              Storage Instructions Keep as concentrated solution. Store at 4°C short term. For extended storage aliquot and store at -20°C or below. Avoid freeze-thaw cycles.
              Storage Buffer PBS

              原創作者:上海篤瑪生物科技有限公司

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