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リコンビナントRabMAb

Anti-GATA1 抗体 [EP2819Y] (ab76121)

Submit a review Q&A (1)
Western blot - Anti-GATA1 antibody [EP2819Y] (ab76121)
  • Western blot - Anti-GATA1 antibody [EP2819Y] (ab76121)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2819Y] to GATA1
  • Suitable for: WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-GATA1 antibody [EP2819Y]
    GATA1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EP2819Y] to GATA1

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WBmore details
    適用なし: ICC/IF,IHC-P or IP

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: K562 and HEL whole cell lysates.

  • 特記事項

    This product has switched from a hybridoma to recombinant production method on 9th June 2023.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

製品の特性

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ab76121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
WB
1/2000. Predicted molecular weight: 43 kDa.
特記事項
WB
1/2000. Predicted molecular weight: 43 kDa.
追加情報
Is unsuitable for ICC/IF,IHC-P or IP.

ターゲット情報

  • 機能

    Transcriptional activator which probably serves as a general switch factor for erythroid development. It binds to DNA sites with the consensus sequence [AT]GATA[AG] within regulatory regions of globin genes and of other genes expressed in erythroid cells.

  • 組織特異性

    Erythrocytes.

  • 関連疾患

    Defects in GATA1 are the cause of X-linked dyserythropoietic anemia and thrombocytopenia (XDAT) [MIM:300367]. XDAT is a disorder characterized by erythrocytes with abnormal size and shape, and paucity of platelets in peripheral blood. The bone marrow contains abundant and abnormally small megakaryocytes.
    Defects in GATA1 are the cause of X-linked thrombocytopenia with beta-thalassemia (XLTT) [MIM:314050]; also knwon as thrombocytopenia, platelet dysfunction, hemolysis, and imbalanced globin synthesis. XLTT consists of an unusual form of thrombocytopenia with beta-thalassemia. Patients have splenomegaly and petechiae, moderate thrombocytopenia, prolonged bleeding time due to platelet dysfunction, reticulocytosis and unbalanced hemoglobin chain synthesis resembling that of beta-thalassemia minor.
    Defects in GATA1 are the cause of anemia without thrombocytopenia X-linked (XLAWT) [MIM:300835]. XLAWT is a form of anemia characterized by abnormal morphology of erythrocytes and granulocytes in peripheral blood, bone marrow dysplasia with hypocellularity of erythroid and granulocytic lineages, and normal or increased number of megakaryocytes. Neutropenia of a variable degree is present in affected individuals.

  • 配列類似性

    Contains 2 GATA-type zinc fingers.

  • ドメイン

    The two fingers are functionally distinct and cooperate to achieve specific, stable DNA binding. The first finger is necessary only for full specificity and stability of binding, whereas the second one is required for binding.

  • 翻訳後修飾

    Highly phosphorylated on serine residues. Phosphorylation on Ser-310 is enhanced on erythroid differentiation. Phosphorylation on Ser-142 promotes sumoylation on Lys-137.
    Sumoylation on Lys-137 is enhanced by phosphorylation on Ser-142 and by interaction with PIAS4. Sumoylation by SUMO1 has no effect on transcriptional activity.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • Anemia, X-linked, without thrombocytopenia, included antibody
      • ERYF 1 antibody
      • Eryf1 antibody
      • Erythroid transcription factor antibody
      • Erythrold transcription factor 1 antibody
      • GATA 1 antibody
      • GATA binding factor 1 antibody
      • GATA binding protein 1 (globin transcription factor 1) antibody
      • GATA binding protein 1 antibody
      • GATA-1 antibody
      • GATA-binding factor 1 antibody
      • GATA1 antibody
      • GATA1_HUMAN antibody
      • GF 1 antibody
      • GF-1 antibody
      • GF1 antibody
      • Globin transcription factor 1 antibody
      • NF E1 antibody
      • NF E1 DNA binding protein antibody
      • NF-E1 DNA-binding protein antibody
      • NFE 1 antibody
      • NFE1 antibody
      • Nuclear factor erythroid 1 antibody
      • Transcription factor GATA1 antibody
      • XLANP antibody
      • XLTDA antibody
      • XLTT antibody
      see all

    画像

    • Western blot - Anti-GATA1 antibody [EP2819Y] (ab76121)
      Western blot - Anti-GATA1 antibody [EP2819Y] (ab76121)
      All lanes : Anti-GATA1 antibody [EP2819Y] (ab76121) at 1/1000 dilution

      Lane 1 : Wild-type K562 cell lysate
      Lane 2 : GATA1 knockout K562 cell lysate
      Lane 3 : HL-60 cell lysate
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 43 kDa



      False colour image of Western blot: Anti-GATA1 antibody [EP2819Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab76121 was shown to bind specifically to GATA1. A band was observed at 47 kDa in wild-type K562 cell lysates with no signal observed at this size in GATA1 knockout cell line ab285360 (knockout cell lysate ab289686). To generate this image, wild-type and GATA1 knockout K562 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    • Western blot - Anti-GATA1 antibody [EP2819Y] (ab76121)
      Western blot - Anti-GATA1 antibody [EP2819Y] (ab76121)
      All lanes : Anti-GATA1 antibody [EP2819Y] (ab76121) at 1/2000 dilution

      Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 2 : HEL (Human Erythroleukemia erythroblast) whole cell lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 43 kDa
      Observed band size: 43 kDa


      Exposure time: 20 seconds


      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    参考文献 (0)

    ab76121 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。

    ab76121 は論文での使用が確認できていません。

    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

    Question

    Inquiry: Dear Sir or Madam, we are interested in doing intracellular GATA1 staining combined with surface marker staining for flow cytometry in human erythroid cells and came across the antibody ab76121 on your webpage. We were wondering whether it comes with a specific protocol, rather than the general protocols supplied on your webpage. We were also going to use K562 cells, which apparently have been tested as shown by the FACS histogramm on the product site. Would it be possible to obtain that protocol, along with the buffers used/recommened and possibly supplied by your company. Also what isotype control (catalog no.) is recommended? Best regards,

    Read More
    Answer

    Thank you for contacting us. The antibody was testing using the protocol below. The rabbit monoclonal isotyope control is ab125938.

    1. Solutions and Reagents

    1.1. 1X PBS

    1.2. Blocking buffer: 0.5% BSA in 1X PBS

    1.3. 2% paraformaldehyde (1% solution - optional for storing samples)

    1.4. 1X FACS permeabilizing solution

    1.5. Fluorescently-conjugated secondary antibody (various forms)

    2. Protocol

    2.1. Collect 1x106 cells/sample.

    2.2. Wash cells once with blocking buffer.

    2.3. Fix cells with 2% paraformaldehyde and incubate at room temperature for 10 min.

    2.4. Wash cells once with blocking buffer.

    2.5. Add 0.5 ml 1X FACS permeabilizing solution and incubate at room temperature for 10 min.

    2.6. Wash cells once with blocking buffer.

    2.7. Incubate cells in blocking buffer for 30 min at room temperature.

    2.8. Add primary antibody 1/50 and incubate for 30 min at room temperature.

    2.9. Wash twice with blocking buffer and incubate with fluorescently-conjugated secondary antibody for 30 min at room temperature.

    2.10. Wash cells twice with blocking buffer.

    2.11. Re-suspend cells in 1X PBS and analyze on flow cytometry. Samples can be kept in 1% paraformaldehyde at 4 °C overnight.

    Read More

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