Anti-phospho-BRCA1 (Ser114) Antibody, clone 3C10G8

Кат. номер

MABC1639-25UL

MABC1639-100UL

Breast cancer type 1 susceptibility protein (UniProt: P38398; also known as EC:2.3.2.27, RING finger protein 53, RING-type E3 ubiquitin transferase BRCA1) is encoded by the BRCA1 (also known as RNF53) gene (Gene ID: 672) in human. BRCA1 and BRCA2 are important in maintaining genomic stability by promoting efficient and precise repair of double-strand breaks. BRCA1 is expressed in all cells and plays a broader role in various cellular processes in response to DNA damage. It promotes the protection of nascent DNA at stalled replication forks independently of the canonical BRCA1-PALB2 interaction. Its recruitment to DNA damage sites is mediated by ABRAXAS1 and the BRCA1-A complex. BRCA1 contains two BRCT domains (aa 1642-1736 and 1756-1855) that recognize and bind phosphorylated pSXXF motif on proteins. The interaction with the phosphorylated pSXXF motif of ABRAXAS1 recruits BRCA1 at DNA damage sites. BRCA1 is reported to exist as a heterodimeric complex with BARD1, a structurally related protein that also contains a N-terminal RING-finger domain and two C-terminal BRCT motifs. This heterodimer regulates the localization and loading of RAD51 at break sites through PALB2-BRCA2. Their association is mediated through their respective RING domains, and together they exhibit E3 ubiquitin ligase activity, which can be disrupted by mutations within the RING domain of BRCA1. BRCA1 and BARD1 are shown to be phosphorylated at residues that are located close to the site of interaction between BARD1 and RAD51 (Ser148 on BARD1 and Ser114 on BRCA1) and Ser114 phosphorylation promotes BRCA1 interaction with PIN1. PIN1 increases interactions between the BRCA1-BARD1 complex and RAD51 resulting in recruitment of RAD51 to stalled replication forks and enhance protection of the nascent DNA strand. Cells lacking BRCA1 show defects in DNA repair by homologous recombination. Mutations in BRCA1 gene have been linked to breast and ovarian cancers. A vast majority of inherited breast cancers result from mutations in the BRCA 1 gene. Male subjects with BRCA1 mutations exhibit about 3-fold increase in risk of prostate cancer. (Ref.: Daza-Martin, M., et al. (2019). Nature. 571(7766); 521-527; Gudumundsdottir, K., and Ashworth, A. (2006). Oncogene 25; 5864-5874).
Specificity
Clone 3C10G8 is a mouse monoclonal antibody that specifically detects BRCA1 phosphorylated on serine 114.
KLH-conjugated linear peptide corresponding to 14 amino acids surrounding phosphoserine 114 from the N-terminal region of human BRCA1.
Quality Control Testing
Evaluated by Western Blotting with a construct containing GST-tagged recombinant fragment of phosphorylated BRCA1.
Western Blotting Analysis (WB): A 1:500 dilution of this antibody detected a construct containing GST-tagged recombinant BRCA1 fragment phosphorylated on Serine 114 but did not react with non-phosphorylated construct. (BRCA1 construct: Courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine).
Tested Applicaitons
Western Blotting Analysis: A representative lot detected phospho-BRCA1 (Ser114) in Western Blotting applications (Daza-Martin, M., et al. (2019). Nature.;571(7766):521-527).
Immunocytochemistry Analysis: A representative lot detected phospho-BRCA1 (Ser114) in Immunocytochemistry applications (Daza-Martin, M., et al. (2019). Nature.;571(7766):521-527).
Immunofluorescence Analysis: A representative lot detected phospho-BRCA1 (Ser114) in Immunofluorescence applications (Daza-Martin, M., et al. (2019). Nature.;571(7766):521-527).
Immunoprecipitation Analysis: A representative lot immunoprecipitated phospho-BRCA1 (Ser114) in Immunoprecipitation applications (Daza-Martin, M., et al. (2019). Nature.;571(7766):521-527).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Anti-phospho-BRCA1 (Ser114), clone 3C10G8, Cat. No. MABC1639, is a mouse monoclonal antibody that detects BRCA1 phosphorylated on serine 114 and is tested in Immunocytochemistry, Immunofluorescence, Immunoprecipitation, and Western Blotting.
Physical form
Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Storage and Stability
Recommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws.
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
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.

Quality Level	200
biological source	mouse
antibody form	purified antibody
antibody product type	primary antibodies
clone	3C10G8, monoclonal
mol wt	calculated mol wt 207.72 kDa","observed mol wt ~30 kDa
purified by	using protein G
species reactivity	human
species reactivity (predicted by homology)	monkey
packaging	antibody small pack of 100 µL
application(s)	immunocytochemistry: suitable","immunofluorescence: suitable","immunoprecipitation (IP): suitable","western blot: suitable
isotype	IgG1
epitope sequence	N-terminal
conjugate	unconjugated
Protein ID accession no.	NP_009255,
UniProt accession no.	P38398,
shipped in	2-8°C
Gene Information	human ... BRCA1(672)

WGK Germany	WGK 1
Flash Point F	Not applicable
Flash Point C	Not applicable