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Immunofluorescence analysis of BRCA-1 was done on 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with BRCA1 (MS13) Mouse Monoclonal Antibody (MA1138) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear and cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant full length human BRCA1|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:200|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1.5-2.5 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Antibody binding site (i.e.epitope) has been mapped within the N-terminal (1-304 amino acids) of Human BRCA1.
This gene encodes a nuclear phosphoprotein that plays a role in maintaining genomic stability, and it also acts as a tumor suppressor. The encoded protein combines with other tumor suppressors, DNA damage sensors, and signal transducers to form a large multi-subunit protein complex known as the BRCA1-associated genome surveillance complex (BASC). This gene product associates with RNA polymerase II, and through the C-terminal domain, also interacts with histone deacetylase complexes. This protein thus plays a role in transcription, DNA repair of double-stranded breaks, and recombination. Mutations in this gene are responsible for approximately 40% of inherited breast cancers and more than 80% of inherited breast and ovarian cancers. Alternative splicing plays a role in modulating the subcellular localization and physiological function of this gene. Many alternatively spliced transcript variants, some of which are disease-associated mutations, have been described for this gene, but the full-length natures of only some of these variants has been described. A related pseudogene, which is also located on chromosome 17, has been identified.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
BRCA1/BRCA2-containing complex, subunit 1; BRCC1; Breast And Ovarian Cancer Susceptibility Protein 1; breast and ovarian cancer sususceptibility protein 1; breast cancer 1; Breast Cancer 1 Early Onset; breast cancer 1, early onset; BROVCA12; early onset breast cancer 1; Fanconi anemia, complementation group S; IRIS; protein phosphatase 1, regulatory subunit 53; PSCP; RING Finger Protein 53; RNF53; truncated BRCA1; truncated breast and ovarian cancer susceptibility protein 1; truncated breast and ovarian cancer sususceptibility protein 1; truncated breast cancer type 1 susceptibility protein
BRCA1; BRCAI; BRCC1; BROVCA1; FANCS; IRIS; PNCA4; PPP1R53; PSCP; RNF53