Product: BRCA1 Antibody
Catalog: AF6289
Description: Rabbit polyclonal antibody to BRCA1
Application: WB IHC IF/ICC
Reactivity: Human
Prediction: Pig
Mol.Wt.: 220kDa; 208kD(Calculated).
Uniprot: P38398
RRID: AB_2835139

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human
Prediction:
Pig(82%)
Clonality:
Polyclonal
Specificity:
BRCA1 Antibody detects endogenous levels of total BRCA1.
RRID:
AB_2835139
Cite Format: Affinity Biosciences Cat# AF6289, RRID:AB_2835139.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

BRCA 1; BRCA1; BRCA1 DNA repair associated; BRCA1/BRCA2 containing complex subunit 1; BRCA1/BRCA2-containing complex, subunit 1; BRCA1_HUMAN; BRCAI; BRCC 1; BRCC1; Breast and ovarian cancer susceptibility protein 1; Breast Cancer 1; Breast Cancer 1 Early Onset; Breast cancer type 1 susceptibility protein; BROVCA1; FANCS; IRIS; PNCA4; PPP1R53; Protein phosphatase 1 regulatory subunit 53; PSCP; RING finger protein 53; RNF53;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P38398 BRCA1_HUMAN:

Isoform 1 and isoform 3 are widely expressed. Isoform 3 is reduced or absent in several breast and ovarian cancer cell lines.

Description:
This gene encodes a nuclear phosphoprotein that plays a role in maintaining genomic stability and 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 BASC for BRCA1-associated genome surveillance complex.
Sequence:
MDLSALRVEEVQNVINAMQKILECPICLELIKEPVSTKCDHIFCKFCMLKLLNQKKGPSQCPLCKNDITKRSLQESTRFSQLVEELLKIICAFQLDTGLEYANSYNFAKKENNSPEHLKDEVSIIQSMGYRNRAKRLLQSEPENPSLQETSLSVQLSNLGTVRTLRTKQRIQPQKTSVYIELGSDSSEDTVNKATYCSVGDQELLQITPQGTRDEISLDSAKKAACEFSETDVTNTEHHQPSNNDLNTTEKRAAERHPEKYQGSSVSNLHVEPCGTNTHASSLQHENSSLLLTKDRMNVEKAEFCNKSKQPGLARSQHNRWAGSKETCNDRRTPSTEKKVDLNADPLCERKEWNKQKLPCSENPRDTEDVPWITLNSSIQKVNEWFSRSDELLGSDDSHDGESESNAKVADVLDVLNEVDEYSGSSEKIDLLASDPHEALICKSERVHSKSVESNIEDKIFGKTYRKKASLPNLSHVTENLIIGAFVTEPQIIQERPLTNKLKRKRRPTSGLHPEDFIKKADLAVQKTPEMINQGTNQTEQNGQVMNITNSGHENKTKGDSIQNEKNPNPIESLEKESAFKTKAEPISSSISNMELELNIHNSKAPKKNRLRRKSSTRHIHALELVVSRNLSPPNCTELQIDSCSSSEEIKKKKYNQMPVRHSRNLQLMEGKEPATGAKKSNKPNEQTSKRHDSDTFPELKLTNAPGSFTKCSNTSELKEFVNPSLPREEKEEKLETVKVSNNAEDPKDLMLSGERVLQTERSVESSSISLVPGTDYGTQESISLLEVSTLGKAKTEPNKCVSQCAAFENPKGLIHGCSKDNRNDTEGFKYPLGHEVNHSRETSIEMEESELDAQYLQNTFKVSKRQSFAPFSNPGNAEEECATFSAHSGSLKKQSPKVTFECEQKEENQGKNESNIKPVQTVNITAGFPVVGQKDKPVDNAKCSIKGGSRFCLSSQFRGNETGLITPNKHGLLQNPYRIPPLFPIKSFVKTKCKKNLLEENFEEHSMSPEREMGNENIPSTVSTISRNNIRENVFKEASSSNINEVGSSTNEVGSSINEIGSSDENIQAELGRNRGPKLNAMLRLGVLQPEVYKQSLPGSNCKHPEIKKQEYEEVVQTVNTDFSPYLISDNLEQPMGSSHASQVCSETPDDLLDDGEIKEDTSFAENDIKESSAVFSKSVQKGELSRSPSPFTHTHLAQGYRRGAKKLESSEENLSSEDEELPCFQHLLFGKVNNIPSQSTRHSTVATECLSKNTEENLLSLKNSLNDCSNQVILAKASQEHHLSEETKCSASLFSSQCSELEDLTANTNTQDPFLIGSSKQMRHQSESQGVGLSDKELVSDDEERGTGLEENNQEEQSMDSNLGEAASGCESETSVSEDCSGLSSQSDILTTQQRDTMQHNLIKLQQEMAELEAVLEQHGSQPSNSYPSIISDSSALEDLRNPEQSTSEKAVLTSQKSSEYPISQNPEGLSADKFEVSADSSTSKNKEPGVERSSPSKCPSLDDRWYMHSCSGSLQNRNYPSQEELIKVVDVEEQQLEESGPHDLTETSYLPRQDLEGTPYLESGISLFSDDPESDPSEDRAPESARVGNIPSSTSALKVPQLKVAESAQSPAAAHTTDTAGYNAMEESVSREKPELTASTERVNKRMSMVVSGLTPEEFMLVYKFARKHHITLTNLITEETTHVVMKTDAEFVCERTLKYFLGIAGGKWVVSYFWVTQSIKERKMLNEHDFEVRGDVVNGRNHQGPKRARESQDRKIFRGLEICCYGPFTNMPTDQLEWMVQLCGASVVKELSSFTLGTGVHPIVVVQPDAWTEDNGFHAIGQMCEAPVVTREWVLDSVALYQCQELDTYLIPQIPHSHY

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
82
Bovine
70
Dog
64
Horse
0
Sheep
0
Xenopus
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P38398 As Substrate

Site PTM Type Enzyme
Sumoylation
M1 Acetylation
K32 Ubiquitination
S36 Phosphorylation
K38 Ubiquitination
K50 Acetylation
K56 Ubiquitination
K65 Ubiquitination
K109 Sumoylation
S114 Phosphorylation
K119 Sumoylation
S123 Phosphorylation
T164 Phosphorylation
S184 Phosphorylation
S186 Phosphorylation
S187 Phosphorylation
T208 Phosphorylation
S220 Phosphorylation
K223 Ubiquitination
K294 Ubiquitination
S308 Phosphorylation O14965 (AURKA)
K309 Ubiquitination
K339 Sumoylation
K357 Ubiquitination
S395 Phosphorylation
S398 Phosphorylation
S403 Phosphorylation
S405 Phosphorylation
S423 Phosphorylation
S425 Phosphorylation
S426 Phosphorylation
K428 Ubiquitination
S434 Phosphorylation
K443 Sumoylation
K443 Ubiquitination
K450 Ubiquitination
S451 Phosphorylation
S454 Phosphorylation
K459 Sumoylation
K459 Ubiquitination
K463 Ubiquitination
T509 Phosphorylation P31749 (AKT1)
S510 Phosphorylation
K519 Ubiquitination
S551 Phosphorylation
K566 Ubiquitination
K576 Ubiquitination
K583 Sumoylation
S615 Phosphorylation
S616 Phosphorylation
T617 Phosphorylation
S632 Phosphorylation P11802 (CDK4)
S647 Phosphorylation
K654 Sumoylation
S694 Phosphorylation P31749 (AKT1)
K701 Ubiquitination
S708 Phosphorylation
K711 Ubiquitination
K734 Sumoylation
K739 Sumoylation
S741 Phosphorylation
K748 Ubiquitination
S753 Phosphorylation
S803 Phosphorylation
K820 Ubiquitination
K830 Ubiquitination
S868 Phosphorylation
K918 Sumoylation
T967 Phosphorylation
K970 Ubiquitination
Y978 Phosphorylation
K987 Ubiquitination
S988 Phosphorylation Q683Z8 (CHK2) , O96017 (CHEK2)
S1007 Phosphorylation
S1009 Phosphorylation
S1050 Phosphorylation
K1079 Sumoylation
S1101 Phosphorylation
S1143 Phosphorylation Q13535 (ATR)
S1164 Phosphorylation P53350 (PLK1)
K1171 Ubiquitination
S1174 Phosphorylation
S1178 Phosphorylation
K1179 Ubiquitination
S1187 Phosphorylation
S1189 Phosphorylation P06493 (CDK1) , Q13315 (ATM)
S1191 Phosphorylation P06493 (CDK1)
T1194 Phosphorylation
S1211 Phosphorylation
S1212 Phosphorylation
S1217 Phosphorylation
S1218 Phosphorylation
S1239 Phosphorylation
S1245 Phosphorylation
S1253 Phosphorylation
K1254 Ubiquitination
K1264 Ubiquitination
S1266 Phosphorylation
S1271 Phosphorylation
K1278 Ubiquitination
S1280 Phosphorylation Q13535 (ATR)
S1286 Phosphorylation
T1289 Phosphorylation
S1298 Phosphorylation Q13535 (ATR)
S1328 Phosphorylation
S1330 Phosphorylation Q13315 (ATM)
S1336 Phosphorylation
S1342 Phosphorylation
S1377 Phosphorylation
S1387 Phosphorylation Q13315 (ATM) , Q13535 (ATR)
T1394 Phosphorylation Q13535 (ATR)
S1423 Phosphorylation Q13535 (ATR) , Q13315 (ATM)
S1457 Phosphorylation Q13315 (ATM) , Q13535 (ATR)
S1460 Phosphorylation
S1466 Phosphorylation Q13315 (ATM) , Q13535 (ATR)
S1473 Phosphorylation
K1476 Ubiquitination
S1480 Phosphorylation
S1483 Phosphorylation
S1496 Phosphorylation
S1497 Phosphorylation P06493 (CDK1) , P24941 (CDK2) , Q13315 (ATM)
S1499 Phosphorylation
S1503 Phosphorylation
S1514 Phosphorylation
S1524 Phosphorylation Q13315 (ATM) , Q13535 (ATR)
S1542 Phosphorylation Q13315 (ATM)
S1547 Phosphorylation
T1548 Phosphorylation
T1550 Phosphorylation
Y1552 Phosphorylation P42685 (FRK)
S1572 Phosphorylation P68400 (CSNK2A1)
S1577 Phosphorylation
S1613 Phosphorylation
T1619 Phosphorylation
K1636 Ubiquitination
S1642 Phosphorylation
K1667 Ubiquitination
T1681 Phosphorylation
T1700 Phosphorylation
T1720 Phosphorylation
T1777 Phosphorylation
S1790 Phosphorylation

Research Backgrounds

Function:

E3 ubiquitin-protein ligase that specifically mediates the formation of 'Lys-6'-linked polyubiquitin chains and plays a central role in DNA repair by facilitating cellular responses to DNA damage. It is unclear whether it also mediates the formation of other types of polyubiquitin chains. The E3 ubiquitin-protein ligase activity is required for its tumor suppressor function. The BRCA1-BARD1 heterodimer coordinates a diverse range of cellular pathways such as DNA damage repair, ubiquitination and transcriptional regulation to maintain genomic stability. Regulates centrosomal microtubule nucleation. Required for normal cell cycle progression from G2 to mitosis. Required for appropriate cell cycle arrests after ionizing irradiation in both the S-phase and the G2 phase of the cell cycle. Involved in transcriptional regulation of P21 in response to DNA damage. Required for FANCD2 targeting to sites of DNA damage. May function as a transcriptional regulator. Inhibits lipid synthesis by binding to inactive phosphorylated ACACA and preventing its dephosphorylation. Contributes to homologous recombination repair (HRR) via its direct interaction with PALB2, fine-tunes recombinational repair partly through its modulatory role in the PALB2-dependent loading of BRCA2-RAD51 repair machinery at DNA breaks. Component of the BRCA1-RBBP8 complex which regulates CHEK1 activation and controls cell cycle G2/M checkpoints on DNA damage via BRCA1-mediated ubiquitination of RBBP8. Acts as a transcriptional activator.

PTMs:

Phosphorylation at Ser-308 by AURKA is required for normal cell cycle progression from G2 to mitosis. Phosphorylated in response to IR, UV, and various stimuli that cause checkpoint activation, probably by ATM or ATR. Phosphorylation at Ser-988 by CHEK2 regulates mitotic spindle assembly.

Autoubiquitinated, undergoes 'Lys-6'-linked polyubiquitination. 'Lys-6'-linked polyubiquitination does not promote degradation.

Subcellular Location:

Nucleus. Chromosome. Cytoplasm.
Note: Localizes at sites of DNA damage at double-strand breaks (DSBs); recruitment to DNA damage sites is mediated by ABRAXAS1 and the BRCA1-A complex (PubMed:26778126). Translocated to the cytoplasm during UV-induced apoptosis (PubMed:20160719).

Cytoplasm.

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Isoform 1 and isoform 3 are widely expressed. Isoform 3 is reduced or absent in several breast and ovarian cancer cell lines.

Subunit Structure:

Heterodimer with BARD1. Part of the BRCA1-associated genome surveillance complex (BASC), which contains BRCA1, MSH2, MSH6, MLH1, ATM, BLM, PMS2 and the MRE11-RAD50-NBN protein (MRN) 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 BRCA1, BARD1, UIMC1/RAP80, ABRAXAS1, BRCC3/BRCC36, BABAM2 and BABAM1/NBA1. Interacts (via the BRCT domains) with ABRAXAS1 (phosphorylated form); this is important for recruitment to sites of DNA damage. Can form a heterotetramer with two molecules of ABRAXAS1 (phosphorylated form). Component of the BRCA1-RBBP8 complex. Interacts (via the BRCT domains) with RBBP8 ('Ser-327' phosphorylated form); the interaction ubiquitinates RBBP8, regulates CHEK1 activation, and involves RBBP8 in BRCA1-dependent G2/M checkpoint control on DNA damage. Associates with RNA polymerase II holoenzyme. Interacts with SMC1A, NELFB, DCLRE1C, CLSPN. Interacts with CHEK1, CHEK2, BAP1, BRCC3, AURKA, UBXN1 and PCLAF. Interacts (via BRCT domains) with BRIP1 (phosphorylated form). Interacts with FANCD2 (ubiquitinated form). Interacts with H2AX (phosphorylated on 'Ser-140'). Interacts (via the BRCT domains) with ACACA (phosphorylated form); the interaction prevents dephosphorylation of ACACA. Part of a BRCA complex containing BRCA1, BRCA2 and PALB2. Interacts directly with PALB2; the interaction is essential for its function in HRR. Interacts directly with BRCA2; the interaction occurs only in the presence of PALB2 which serves as the bridging protein. Interacts (via the BRCT domains) with LMO4; the interaction represses the transcriptional activity of BRCA1. Interacts (via the BRCT domains) with CCAR2 (via N-terminus); the interaction represses the transcriptional activator activity of BRCA1. Interacts with EXD2. Interacts (via C-terminus) with DHX9; this interaction is direct and links BRCA1 to the RNA polymerase II holoenzyme.

Family&Domains:

The BRCT domains recognize and bind phosphorylated pSXXF motif on proteins. The interaction with the phosphorylated pSXXF motif of ABRAXAS1, recruits BRCA1 at DNA damage sites.

The RING-type zinc finger domain interacts with BAP1.

Research Fields

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Genetic Information Processing > Replication and repair > Homologous recombination.

· Genetic Information Processing > Replication and repair > Fanconi anemia pathway.

· Genetic Information Processing > Folding, sorting and degradation > Ubiquitin mediated proteolysis.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

References

1). Li et al. PARP1 Inhibitor Combined With Oxaliplatin Efficiently Suppresses Oxaliplatin Resistance in Gastric Cancer-Derived Organoids via Homologous Recombination and the Base Excision Repair Pathway. Frontiers in Cell and Developmental Biology [IF=5.5]

Application: WB    Species: Human    Sample:

FIGURE 6. Treatment of Oxaliplatin inhibits HR repair pathways via blocking CDK1-BRCA1 activities in Oxaliplatin resistance cell line. (A) Verification by WB on the effects of Olaparib + Oxaliplatin, Oxaliplatin, Olaparib, AG-02432 and cisplatin on CDK1 expression and its phosphorylation, BRCA1 expression and its phosphorylation, RAD51 expression in SNU719, MKN74, and AGS Oxaliplatin resistance strains. Drug action time was 36 h. (B) Histochemical results of protein phosphorylation in gastric cancer patients. (C,D) The effects of Olaparib + Oxaliplatin and cisplatin combined with CDK1 inhibitor Olaparib on colony formation of overexpressed PARP1 and normally expressed PARP1 cell lines in SNU719, MKN74, and AGS Oxaliplatin resistance strains. Colonies were stained with crystal violet. The Student’s t test was used for statistical analysis. Error bars indicate mean ± standard deviation. OXA, Oxaliplatin. OLP, Olaparib. CON, control group. CISP, cisplatin. PCDK1, CDK1 phosphorylation antibody. PBRCA1, BRCA1 phosphorylation antibody. AGSR, AGS Oxaliplatin resistance. SNU719R, SNU719 Oxaliplatin resistance. MKN74R, MKN74 Oxaliplatin resistance. ∗< 0.05. All experiments were repeated three times.

2). Xiao M et al. Let-7e Suppresses DNA Damage Repair and Sensitizes Ovarian Cancer to Cisplatin through Targeting PARP1. MOLECULAR CANCER RESEARCH 2019 Nov 13 (PubMed: 31722968) [IF=5.2]

Application: WB    Species: human    Sample: ovarian

Fig. 3. |PARP1 promotes DNA DSB repair in ovarian cancer.D and E, Expression of PARP1, BRCA1, and Rad51 in conditions where shPARP1 or PARP1 cDNA were transfected. Three experiments were done.

3). Ye Y et al. Anticancer Effect of Puerarin on Ovarian Cancer Progression Contributes to the Tumor Suppressor Gene Expression and Gut Microbiota Modulation. Journal of Immunology Research 2022 Jul 28;2022:4472509. (PubMed: 35935578) [IF=4.1]

Application: WB    Species: Rat    Sample: tumor tissues

Figure 5 Expression of cell proliferation and tumor suppressor genes was regulated by puerarin treatment in OC model rats. (a) Positive expression of Ki67, PCNA, P53, and PTEN in tumor tissues was detected by an immunohistochemistry assay (magnification: ×200). (b) Protein expression levels of P53, P21, PTEN, BRCA1, BIRC5, and CTGF in tumor tissues were detected by Western blot assay, and GAPDH was used as the internal control. OC: ovarian cancer.

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