Overview

  • Product nameAnti-BRCA1 antibody [MS110]
    See all BRCA1 primary antibodies
  • Description
    Mouse monoclonal [MS110] to BRCA1
  • Tested applicationsSuitable for: Flow Cyt, ICC/IF, IHC-Fr, IP, IHC-Pmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein (Human).

  • EpitopeWithin the N-terminal 304 amino acids of BRCA1.
  • Positive control
    • This antibody gave a positive signal in human breast carcinoma formalin-fixed, paraffin embedded tissue sections.
  • General notes

    Please note that this antibody is not suitable for WB.
    Despite positive publications and Abreviews we have mixed feedback on this antibody in WB and we do not guarantee ab16780 for WB.

Properties

Applications

Our Abpromise guarantee covers the use of ab16780 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notesIs unsuitable for WB.
  • Target

    • FunctionE3 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.
    • Tissue specificityIsoform 1 and isoform 3 are widely expressed. Isoform 3 is reduced or absent in several breast and ovarian cancer cell lines.
    • PathwayProtein modification; protein ubiquitination.
    • Involvement in diseaseDefects in BRCA1 are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case. Note=Mutations in BRCA1 are thought to be responsible for 45% of inherited breast cancer. Moreover, BRCA1 carriers have a 4-fold increased risk of colon cancer, whereas male carriers face a 3-fold increased risk of prostate cancer. Cells lacking BRCA1 show defects in DNA repair by homologous recombination.
      Defects in BRCA1 are a cause of susceptibility to breast-ovarian cancer familial type 1 (BROVCA1) [MIM:604370]. A condition associated with familial predisposition to cancer of the breast and ovaries. Characteristic features in affected families are an early age of onset of breast cancer (often before age 50), increased chance of bilateral cancers (cancer that develop in both breasts, or both ovaries, independently), frequent occurrence of breast cancer among men, increased incidence of tumors of other specific organs, such as the prostate. Note=Mutations in BRCA1 are thought to be responsible for more than 80% of inherited breast-ovarian cancer.
      Defects in BRCA1 are a cause of genetic susceptibility to ovarian cancer [MIM:113705].
    • Sequence similaritiesContains 2 BRCT domains.
      Contains 1 RING-type zinc finger.
    • DomainThe BRCT domains recognize and bind phosphorylated pSXXF motif on proteins. The interaction with the phosphorylated pSXXF motif of FAM175A/Abraxas, recruits BRCA1 at DNA damage sites.
      The RING-type zinc finger domain interacts with BAP1.
    • Post-translational
      modifications
      Phosphorylation at Ser-308 by STK6/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.
      Autoubiquitinated, undergoes 'Lys-6'-linked polyubiquitination. 'Lys-6'-linked polyubiquitination does not promote degradation.
    • Cellular localizationCytoplasm; 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.
    • Information by UniProt
    • Database links
    • Alternative names
      • BRCA 1 antibody
      • BRCA1 antibody
      • BRCA1/BRCA2 containing complex subunit 1 antibody
      • BRCA1/BRCA2-containing complex, subunit 1 antibody
      • BRCA1_HUMAN antibody
      • BRCAI antibody
      • BRCC 1 antibody
      • BRCC1 antibody
      • Breast and ovarian cancer susceptibility protein 1 antibody
      • Breast Cancer 1 antibody
      • Breast Cancer 1 Early Onset antibody
      • Breast cancer type 1 susceptibility protein antibody
      • BROVCA1 antibody
      • IRIS antibody
      • PNCA4 antibody
      • PPP1R53 antibody
      • Protein phosphatase 1 regulatory subunit 53 antibody
      • PSCP antibody
      • RING finger protein 53 antibody
      • RNF53 antibody
      see all

    Anti-BRCA1 antibody [MS110] images

    • ICC/IF image of ab16780 stained MCF7cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16780, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • IHC image of ab16780 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16780, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Overlay histogram showing MCF7 cells stained with ab16780 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16780, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • ab16780 staining BRAC1 in Human Ovarian Tumor cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Acetone:Methanol and blocked with DAKO Protein Block, Serum-Free for 1 hour at 18°C. Samples were incubated with primary antibody (1/100) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated Rabbit anti-mouse IgG (H+L) polyclonal (1/400) was used as the secondary antibody.

      See Abreview

    • ab16780 staining BRCA1 in Human skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in formaldehyde and subjected to heat-mediated antigen retrieval in citrate buffer (pH 6.0) prior to blocking with 2% BSA for 1 hour at 22°C. The primary antibody was diluted 1/50 and incubated with the sample for 20 hours at 4°C. A Biotin-conjugated goat anti-mouse polyclonal was used as the secondary antibody, diluted 1/800. Antibody was detected by DAB staining.

      See Abreview

    • ab16780 staining BRAC1 in Human colon cancer cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 and blocked with 3% BSA for 30 minutes at 4°C. Samples were incubated with primary antibody (1/200) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG (H+L) polyclonal (1/1000) was used as the secondary antibody.

      See Abreview

    • ab16780 staining BRCA1 in human A431 epidermoid cancer cells by ICC/IF (immunocytochemistry/immunofluorescence). Cells were formaldehyde fixed, permeabilized by Triton X-100 and blocked 5% BSA for 30 minutes at room temperature. The sample was incubated with the primary antibody (1/50 in BSA) for 1 hour. An Alexa Fluor 488®-conjugated Goat anti-mouse polyclonal (1/50) was used as the secondary.

      See Abreview

    References for Anti-BRCA1 antibody [MS110] (ab16780)

    This product has been referenced in:
    • Li L  et al. Targeting poly(ADP-ribose) polymerase and the c-Myb-regulated DNA damage response pathway in castration-resistant prostate cancer. Sci Signal 7:ra47 (2014). Read more (PubMed: 24847116) »
    • Muñoz MC  et al. An RNF168 fragment defective for focal accumulation at DNA damage is proficient for inhibition of homologous recombination in BRCA1 deficient cells. Nucleic Acids Res 42:7720-33 (2014). WB . Read more (PubMed: 24829461) »

    See all 15 Publications for this product

    Product Wall

    Application Immunoprecipitation
    Immuno-precipitation step Protein G
    Sample Human Cell lysate - whole cell (Hela cells)
    Specification Hela cells
    Total protein in input 2000 µg
    Username

    Dr. Remi Buisson

    Verified customer

    Submitted Jan 05 2015

    Application Western blot
    Loading amount 10 µg
    Gel Running Conditions Reduced Denaturing (4-12%)
    Sample Human Cell lysate - whole cell (UWB1.289 cells)
    Specification UWB1.289 cells
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Remi Buisson

    Verified customer

    Submitted Nov 17 2014

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (Ovarian Tumor)
    Gel Running Conditions Reduced Denaturing (3-8%)
    Loading amount 30 µg
    Specification Ovarian Tumor
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 18°C
    Username

    Abcam user community

    Verified customer

    Submitted May 05 2014

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 18°C
    Sample Human Cell (Ovarian Tumor)
    Specification Ovarian Tumor
    Permeabilization Yes - Acetone:Methanol
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Apr 02 2014

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (6%)
    Sample Human Cell lysate - whole cell (U2OS osteosarcoma)
    Specification U2OS osteosarcoma
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 25 2014

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (6%)
    Sample Mouse Cell lysate - whole cell (Mouse breast cancer cell line)
    Specification Mouse breast cancer cell line
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 15 2013

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 4°C
    Sample Human Cell (Human colon cancer cell line)
    Specification Human colon cancer cell line
    Permeabilization Yes - 0.3% Triton X-100
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Oct 15 2013

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (MCF-7)
    Loading amount 30 µg
    Specification MCF-7
    Gel Running Conditions Reduced Denaturing (7,5)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Abcam user community

    Verified customer

    Submitted May 07 2013

    Thank you for your email.

    As observed in image the problem is more with the protocol rather than antibody. Please try troubleshooting as follows;

    - Heat lysates with sample buffer for 5-10 minutes at 100C
    - Milk, blocking agent is...

    Read More

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    I have read the details you have kindly provided and have following further questions for better understanding of the problem;

    -...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"