BRCA1 Rabbit pAb

Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (BRCA1) Polyclonal Antibody, Unconjugated (AP94454) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (BRCA1) Polyclonal Antibody, Unconjugated (AP94454) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Blank control (Black line): Mouse spleen(Black). Primary Antibody (green line): Rabbit Anti-BRCA1 antibody (AP94454) Dilution: 3 µg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647 Dilution: 1 µg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 10,000 events was performed.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	IHC-P, IHC-F, IF
Reactivity	Mouse
Host	Rabbit
Clonality	Polyclonal
Calculated MW	199 KDa
Physical State	Liquid
Immunogen	KLH conjugated synthetic peptide derived from mouse BRCA1
Epitope Specificity	64-160/1812
Isotype	IgG
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Cytoplasm; Nucleus. Localizes at sites of DNA damage at double-strand breaks (DSBs) and recruitment to DNA damage sites is mediated by the BRCA1-A complex.
SIMILARITY	Contains 2 BRCT domains. Contains 1 RING-type zinc finger.
SUBUNIT	Heterodimer with BARD1. Part of the BRCA1-associated genome surveillance complex (BASC), which contains BRCA1, MSH2, MSH6, MLH1, ATM, BLM, PMS2 and the RAD50-MRE11-NBN protein complex. This association could be a dynamic process changing throughout the cell cycle and within subnuclear domains. Component of the BRCA1-A complex, at least composed of the BRCA1, BARD1, UIMC1/RAP80, FAM175A/Abraxas, BRCC3/BRCC36, BRE/BRCC45 and BABAM1/NBA1. Interacts (via BRCT domains) with FAM175A/Abraxas and RBBP8. Associates with RNA polymerase II holoenzyme. Interacts with SMC1A and COBRA1/NELFB. Interacts (via BRCT domains) with phosphorylated BRIP1. Interacts with FANCD2 (ubiquitinated). Interacts with BAP1. Interacts with DCLRE1C/Artemis and CLSPN. Interacts with H2AFX (phosphorylated on 'Ser-140'). Interacts with CHEK1 and CHEK2. Interacts with BRCC3. Interacts (via BRCT domains) with ACACA (phosphorylated); the interaction prevents dephosphorylation of ACACA. Interacts with AURKA. Interacts with UBXN1. Part of a trimeric complex containing BRCA1, BRCA2 and PALB2. Interacts with PALB2 and this interaction is essential for its function in HRR. Interacts with BRCA2 only in the presence of PALB2 which serves as the bridging protein.
Post-translational modifications	Phosphorylation at Ser-308 by AURKA is required for normalcell cycle progression from G2 to mitosis. Phosphorylated inresponse to IR, UV, and various stimuli that cause checkpointactivation, probably by ATM or ATR. Phosphorylation at Ser-988 byCHEK2 regulates mitotic spindle assembly.Autoubiquitinated, undergoes 'Lys-6'-linkedpolyubiquitination. 'Lys-6'-linked polyubiquitination does notpromote degradation.
DISEASE	Defects in BRCA1 are a cause of susceptibility to breastcancer (BC) [MIM:114480]. A common malignancy originating frombreast epithelial tissue. Breast neoplasms can be distinguished bytheir histologic pattern. Invasive ductal carcinoma is by far themost common type. Breast cancer is etiologically and geneticallyheterogeneous. Important genetic factors have been indicated byfamilial occurrence and bilateral involvement. Mutations at morethan one locus can be involved in different families or even in thesame case. Note=Mutations in BRCA1 are thought to be responsiblefor 45% of inherited breast cancer. Moreover, BRCA1 carriers have a4-fold increased risk of colon cancer, whereas male carriers face a3-fold increased risk of prostate cancer. Cells lacking BRCA1 showdefects in DNA repair by homologous recombination.Defects in BRCA1 are a cause of susceptibility tofamilial breast-ovarian cancer type 1 (BROVCA1) [MIM:604370]. Acondition associated with familial predisposition to cancer of thebreast and ovaries. Characteristic features in affected familiesare an early age of onset of breast cancer (often before age 50),increased chance of bilateral cancers (cancer that develop in bothbreasts, or both ovaries, independently), frequent occurrence ofbreast cancer among men, increased incidence of tumors of otherspecific organs, such as the prostate. Note=Mutations in BRCA1 arethought to be responsible for more than 80% of inheritedbreast-ovarian cancer.Defects in BRCA1 are a cause of susceptibility to ovariancancer (OC) [MIM:167000]. The term ovarian cancer definesmalignancies originating from ovarian tissue. Although manyhistologic types of ovarian tumors have been described, epithelialovarian carcinoma is the most common form. Ovarian cancers areoften asymptomatic and the recognized signs and symptoms, even oflate-stage disease, are vague. Consequently, most patients arediagnosed with advanced disease.Defects in BRCA1 are a cause of susceptibility topancreatic cancer type 4 (PNCA4)
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	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].

Additional Information
Target/Specificity	Isoform 1 and isoform 3 are widely expressed. Isoform 3 is reduced or absent in several breast and ovarian cancer cell lines.
Dilution	IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=3ug/Test
Format	0.01M TBS(pH7.4) with 1% BSA, 0.09% (W/V) sodium azide and 50% Glyce
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol