Monoclonal Anti-BRCA1 antibody produced in mouse
SIGMA/SAB2702136
Synonym: PNCA4; PPP1R53; PSCP; RNF53
Product Type: Product-on-demand
Immunofluorescence BRCA1 antibody 6B4 (SAB2702136) 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).
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).
Western Blotting 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 (SAB2702136) at a dilution of 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
Western Blotting BRCA1 antibody [6B4] (SAB2702136) was used at 1:1000 dilution for western blot assay of lysates from cells transfected with control or BRCA1-specific siRNA. Lysates were prepared at the indicated times following transfection. RAD50 antibody [13B3] (SAB2702155) was used as a loading control.
Western Blotting BRCA1 antibody 6B4 (SAB2702136) 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.
Western Blotting 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 (SAB2702136) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
Immunoprecipitation BRCA1 antibody 6B4 (SAB2702136) and BRCA1 antibody 17F8 (SAB2702134) 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.
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.
ChIP Assay BRCA1 antibody 6B4 (SAB2702136) and BRCA1 antibody 17F8 (SAB2702134) 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.
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.
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | 6B4 |
| monoclonal | |
| concentration | 1.36 mg/mL |
| conjugate | unconjugated |
| form | liquid |
| isotype | IgG1 |
| mol wt | 220 kDa |
| Quality Level | 100  |
| shipped in | wet ice |
| species reactivity | mouse, human |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | ChIP: suitable |
| immunocytochemistry: suitable | |
| immunofluorescence: suitable | |
| immunohistochemistry: suitable | |
| immunoprecipitation (IP): suitable | |
| western blot: 1:500-1:3000 | |
| UniProt accession no. | P38398  |
| Application: | 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. |
| Biochem/physiol Actions: | BRCA1-6B4 recognizes full length 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. In a high proportion of breast and ovarian cancer cell lines, BRCA1 aberrantly mislocates to the cytoplasm. Recent studies suggest a role for BRCA1 in DNA double strand break repair because of its association with Rad51. BRCA1 is predominantly expressed in proliferating and differentiating cells and contributes to cellular growth. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| Features and Benefits: | Evaluate our antibodies with complete peace of mind. If the antibody does not perform in your application, we will issue a full credit or replacement antibody. Learn more. |
| General description: | The gene BRCA1 is mapped to human chromosome 17q21 and codes for a protein with zinc finger motif and a set of acidic residues. BRCA1 is localized to the nucleus and cytoplasm. |
| Immunogen: | BRCA1 protein fragment expressed in E. coli corresponding to amino acids 341-748. |
| Physical form: | Phosphate-buffered saline, no preservative added. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |