Immunohistochemistry analysis of BRCA1 was performed on human breast carcinoma tissue. To expose target proteins, antigen retrieval was performed by microwaving tissues for 8-15 minutes in 10mM sodium citrate buffer (pH 6.0). Following antigen retrieval, endogenous peroxidases were blocked with 3% hydrogen peroxide-methanol for 15 min at room temperature. Tissue slides were washed with deionized water and PBS, and then blocked in 3% BSA-PBS for 30 min at room temperature. Tissues were probed with a BRCA1 monoclonal antibody (Product # MA1-137) diluted 1:100 in 3% BSA-PBS (right panel) or incubated with buffer alone not containing primary antibody as a negative control (left panel), overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human|
|Published 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)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (Frozen) (IHC (F))||1:50-1:100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
|Western Blot (WB)||See 1 publications below|
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 haploinsufficiency for replication stress suppression in primary cells.
MA1-137 was used in western blot to assess the functionality of BRCA1 in the heterozygous state.
|Pathania S,Bade S,Le Guillou M,Burke K,Reed R,Bowman-Colin C,Su Y,Ting DT,Polyak K,Richardson AL,Feunteun J,Garber JE,Livingston DM||Nature communications (5:null)||2014|