APPLICATION
Application Note
*Optimal dilutions/concentrations should be determined by the researcher.
Application |
Recommended Dilution |
1:500-1:3000 |
1:100-1:1000 |
Assay dependent |
Assay dependent |
Assay dependent |
Assay dependent |
Not tested in other applications.
Calculated MW
Positive Control
293T , MCF7 , U2OS , HeLa
Product Note
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.
PROPERTIES
Form
Liquid
Buffer
PBS, 20% Glycerol
Preservative
No Preservative
Storage
Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.
Concentration
1 mg/ml (Please refer to the vial label for the specific concentration.)
Antigen Species
Human
Immunogen
BRCA1 protein fragment expressed in E. coli corresponding to amino acids 341-748.
Purification
Protein G purified
Conjugation
Unconjugated
RRID
AB_368616
Note
For laboratory research use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Purchasers shall not, and agree not to enable third parties to, analyze, copy, reverse engineer or otherwise attempt to determine the structure or sequence of the product.
TARGET
Synonyms
BRCA1 DNA repair associated , BRCAI , BRCC1 , BROVCA1 , FANCS , IRIS , PNCA4 , PPP1R53 , PSCP , RNF53
Cellular Localization
Nucleus , Cytoplasm
Background
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. [provided by RefSeq, May 2009]
Database
Research Area
DATA IMAGES
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GTX70115 IP Image
BRCA1 antibody 6B4 (GTX70115) and BRCA1 antibody 17F8 (GTX70111) 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.
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GTX70115 WB Image
Wild-type (WT) and BRCA1 knockout (KO) HeLa cell extracts (30 μg) were separated by 5% SDS-PAGE, and the membrane was blotted with BRCA1 antibody [6B4] - ChIP grade (GTX70115) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.
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GTX70115 ChIP assay Image
BRCA1 antibody 6B4 (GTX70115) and BRCA1 antibody 17F8 (GTX70111) 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.
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GTX70115 WB Image
Various whole cell extracts (30 μg) were separated by 5% SDS-PAGE, and the membrane was blotted with BRCA1 antibody [6B4] - ChIP grade (GTX70115) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody, and the signal was developed with Trident femto Western HRP Substrate.
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GTX70115 IHC Image
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GTX70115 WB Image
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 (GTX70115) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody.
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GTX70115 WB Image
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 (GTX70115) at a dilution of 1:500. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody.
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GTX70115 WB Image
BRCA1 antibody [6B4] (GTX70115) 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] (GTX70228) was used as a loading control.
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GTX70115 ICC/IF Image
BRCA1 antibody 6B4 (GTX70115) 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).
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REFERENCE
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