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| Product Name | BRCA1 antibody - ChIP grade |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description |
|
| Immunogen | BRCA1 protein fragment expressed in E. coli corresponding to amino acids 341-748. |
| 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 |
| Gene Synonyms | 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 |
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.
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.
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).
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.
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.
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.: |
| 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 |
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