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    brca1-antibody-ms110-ab16780.pdf

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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Ubiquitylation
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Anti-BRCA1 抗体 [MS110] (ab16780)

  • Datasheet
  • SDS
Reviews (14)Q&A (9)References (56)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)
  • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
  • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
  • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
  • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
  • Flow Cytometry - Anti-BRCA1 antibody [MS110] (ab16780)

Key features and details

  • Mouse monoclonal [MS110] to BRCA1
  • Suitable for: Flow Cyt, ICC/IF, IHC-P
  • Reacts with: Human
  • Isotype: IgG1

こちらの製品もご検討ください

Biochemical
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二次抗体
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Goat Anti-Mouse IgG H&L (HRP) (ab205719)
一次抗体
Product image
Anti-BRCA1 antibody [EPR19433] (ab213929)

関連製品

製品の概要

  • 製品名

    Anti-BRCA1 antibody [MS110]
    BRCA1 一次抗体 製品一覧
  • 製品の詳細

    Mouse monoclonal [MS110] to BRCA1
  • 由来種

    Mouse
  • アプリケーション

    適用あり: Flow Cyt, ICC/IF, IHC-Pmore details
    適用なし: WB
  • 種交差性

    交差種: Human
  • 免疫原

    Recombinant full length protein corresponding to Human BRCA1.

  • エピトープ

    Within the N-terminal 304 amino acids of BRCA1.
  • ポジティブ・コントロール

    • IHC-P: Human breast carcinoma tissue. Human skin tissue. ICC/IF: MCF7 and A431 cells. Human ovarian tumor cells. Human colon cancer cells. Flow cytometry: MCF7 cells.
  • 特記事項

    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.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • 精製度

    Protein G purified
  • ポリ/モノ

    モノクローナル
  • クローン名

    MS110
  • ミエローマ

    NS1
  • アイソタイプ

    IgG1
  • 軽鎖の種類

    kappa
  • 研究分野

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Ubiquitylation
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • BRCA1
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E3 Enzymes
    • RING Finger E3 Ligase
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Tumor Suppressors
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other

関連製品

  • Alternative Versions

    • Anti-BRCA1 antibody [MS110] - BSA and Azide free (ab264518)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human BRCA1 protein (ab82204)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab16780の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF (4)
Use at an assay dependent concentration.
IHC-P (2)
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
特記事項
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-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
追加情報
Is unsuitable for WB.

ターゲット情報

  • 機能

    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.
  • 組織特異性

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

    Protein modification; protein ubiquitination.
  • 関連疾患

    Defects 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].
  • 配列類似性

    Contains 2 BRCT domains.
    Contains 1 RING-type zinc finger.
  • ドメイン

    The 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.
  • 翻訳後修飾

    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.
  • 細胞内局在

    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.
  • Target information above from: UniProt accession P38398 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 672 Human
    • Omim: 113705 Human
    • SwissProt: P38398 Human
    • Unigene: 194143 Human
    • 別名

      • BRCA 1 antibody
      • BRCA1 antibody
      • BRCA1 DNA repair associated 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
      • FANCS 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

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)

      IHC image of ab16780 staining in normal human breast 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, 1µg/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.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)This image is courtesy of an Anonymous 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

    • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
      Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)

      ICC/IF image of ab16780 stained MCF7cells. The cells were 4% PFA fixed (10 minutes) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16780, 5 µg/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 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

    • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
      Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)Image is courtesy of an anonymous Abreview

      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 a 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

    • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
      Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)Image is courtesy of an anonymous 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

    • Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)
      Immunocytochemistry/ Immunofluorescence - Anti-BRCA1 antibody [MS110] (ab16780)This image is courtesy of an Abreview submitted by Dr Alejandro Vazquez-Martin
      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

    • Flow Cytometry - Anti-BRCA1 antibody [MS110] (ab16780)
      Flow Cytometry - Anti-BRCA1 antibody [MS110] (ab16780)

      Overlay histogram showing MCF7 cells stained with ab16780 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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, 1 µg/1x106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 µg/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 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.

    プロトコール

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    データシートおよび資料

    • SDS download

    • Datasheet download

      Download

    参考文献 (56)

    ab16780 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab16780 は 56 報の論文で使用されています。

    • Patel PS  et al. RNF168 regulates R-loop resolution and genomic stability in BRCA1/2-deficient tumors. J Clin Invest 131:N/A (2021). PubMed: 33529165
    • Minten EV  et al. SIRT2 promotes BRCA1-BARD1 heterodimerization through deacetylation. Cell Rep 34:108921 (2021). PubMed: 33789098
    • McMahon KA  et al. Cavin3 released from caveolae interacts with BRCA1 to regulate the cellular stress response. Elife 10:N/A (2021). PubMed: 34142659
    • Gomez Godinez V  et al. DNA damage induced during mitosis undergoes DNA repair synthesis. PLoS One 15:e0227849 (2020). PubMed: 32343690
    • Somri-Gannam L  et al. IGF1R Axis Inhibition Restores Dendritic Cell Antitumor Response in Ovarian Cancer. Transl Oncol 13:100790 (2020). PubMed: 32428851
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 23 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BRCA1 antibody [MS110]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (HT29 tumor taken out Balb/C nude mice)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Ph6
    Permeabilization
    No
    Specification
    HT29 tumor taken out Balb/C nude mice
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 20 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-BRCA1 antibody [MS110]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Vascular smooth muscle cell)
    Permeabilization
    Yes - NP40
    Specification
    Vascular smooth muscle cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 26 2018

    Western blot abreview for Anti-BRCA1 antibody [MS110]

    Below Average
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Endothelial)
    Gel Running Conditions
    Reduced Denaturing (7%)
    Loading amount
    50 µg
    Specification
    Endothelial
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 05 2017

    Immunoprecipitation abreview for Anti-BRCA1 antibody [MS110]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein G
    Sample
    Human Cell lysate - whole cell (Hela cells)
    Specification
    Hela cells
    Total protein in input
    2000 µg
    Read More

    DR. Remi Buisson

    Verified customer

    投稿 Jan 05 2015

    Western blot abreview for Anti-BRCA1 antibody [MS110]

    Average
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    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
    Read More

    DR. Remi Buisson

    Verified customer

    投稿 Nov 17 2014

    Western blot abreview for Anti-BRCA1 antibody [MS110]

    Below Average
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    投稿 May 05 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-BRCA1 antibody [MS110]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 02 2014

    Western blot abreview for Anti-BRCA1 antibody [MS110]

    Average
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    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
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    Abcam user community

    Verified customer

    投稿 Feb 25 2014

    Western blot abreview for Anti-BRCA1 antibody [MS110]

    Good
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 15 2013

    Question

    Lisates were prepaired in ice using RIPA buffer, from human lung cancer cell lines: SKLMS, CALU-1, A549; The dilution of primary antibody was 1:500; Secondary antibody was used anti-mouse from LSbio, lysates were heated during 1 minute. Unfortunately I still don't have an order number from secretary.
    I've attached the most representative image on this e-mail.
    Thank you for your time,
    Sincerely,

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    Abcam community

    Verified customer

    Asked on Sep 28 2012

    Answer

    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 not recommend for phospho-specific antibody e.g. ab47276; please try 5% BSA
    - ab16780 is mouse monoclonal antibody and ab47276 is rabbit polyclonal. Anti mouse antibody would not be working with ab47276.
    - Please try 4X5 minutes and 1, 10 minutes wash in TBST buffer.
    - It seems the lysates are loaded at high amount. If lysates concentration is correct then I would recommend trying 10ug lysates.

    I hope the suggestions will help to improve results.

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    Abcam Scientific Support

    Answered on Sep 28 2012

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