Anti-RACGAP1/MGCRACGAP 抗体 (ab2270)

Reviews (4)Q&A (2)References (21)

Key features and details

  • Goat polyclonal to RACGAP1/MGCRACGAP
  • Suitable for: ICC/IF, IHC-P, WB, IP
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-RACGAP1/MGCRACGAP antibody
    RACGAP1/MGCRACGAP 一次抗体 製品一覧

  • 製品の詳細

    Goat polyclonal to RACGAP1/MGCRACGAP

  • 由来種

    Goat

  • アプリケーション

    適用あり: ICC/IF, IHC-P, WB, IPmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Cow, Dog

  • 免疫原

    Synthetic peptide corresponding to Human RACGAP1/MGCRACGAP aa 620-632 (C terminal).
    Sequence:

    C-GRQGNFFASPMLK


    (Peptide available as ab22898)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • Human testis lysate. K562 cells.

  • 特記事項

    Principal Names - RACGAP1/MGCRACGAP; Rac GTPase activating protein 1; ID-GAP; MgcRacGAP; GTPase activating protein. Official Gene Symbol - - RACGAP1/MGCRACGAP. GenBank Accession Number – NP_037409. LocusLink ID - 29127.

     This product was previously labelled as RACGAP1

     

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • バッファー

    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...

  • 精製度

    Immunogen affinity purified

  • 特記事項（精製）

    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab2270 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ICC/IF Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml.
WB Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 71 kDa).

1 hour primary incubation is recommended for this product.
IP Use at an assay dependent concentration.

ターゲット情報

  • 機能

    Component of the centralspindlin complex that serves as a microtubule-dependent and Rho-mediated signaling required for the myosin contractile ring formation during the cell cycle cytokinesis. Plays key roles in controlling cell growth and differentiation of hematopoietic cells through mechanisms other than regulating Rac GTPase activity. Also involved in the regulation of growth-related processes in adipocytes and myoblasts. May be involved in regulating spermatogenesis and in the RACGAP1 pathway in neuronal proliferation. Shows strong GAP (GTPase activation) activity towards CDC42 and RAC1 and less towards RHOA. Essential for the early stages of embryogenesis. May play a role in regulating cortical activity through RHOA during cytokinesis. May participate in the regulation of sulfate transport in male germ cells.

  • 組織特異性

    Highly expressed in testis, thymus and placenta. Expressed at lower levels in spleen and peripheral blood lymphocytes. In testis, expression is restricted to germ cells with the highest levels of expression found in spermatocytes. Expression is regulated in a cell cycle-dependent manner and peaks during G2/M phase.

  • 配列類似性

    Contains 1 phorbol-ester/DAG-type zinc finger.
    Contains 1 Rho-GAP domain.

  • ドメイン

    The coiled coil region is indispensible for localization to the midbody during cytokinesis.

  • 翻訳後修飾

    Phosphorylated at multiple sites in the midbody during cytokinesis. Phosphorylation by AURKB on Ser-387 at the midbody is, at least in part, responsible for exerting its latent GAP activity towards RhoA. Phosphorylation on multiple serine residues by PLK1 enhances its association with ECT2 and is critical for cleavage furrow formation.

  • 細胞内局在

    Nucleus. Cytoplasm. Cytoplasm > cytoskeleton > spindle. Cytoplasmic vesicle > secretory vesicle > acrosome. Cleavage furrow. Midbody. Colocalizes with RND2 in Golgi-derived proacrosomal vesicles and the acrosome (By similarity). During interphase, localized to the nucleus and cytoplasm along with microtubules, in anaphase, is redistributed to the central spindle and, in telophase and cytokinesis, to the midbody. Colocalizes with RHOA at the myosin contractile ring during cytokinesis. Colocalizes with ECT2 to the mitotic spindles during anaphase/metaphase, the cleavage furrow during telophase and at the midbody at the end of cytokinesis. Colocalizes with Cdc42 to spindle microtubules from prometaphase to telophase.
  • Information by UniProt

  • 参照データベース

    • 別名

      • CYK4 antibody
      • GAP antibody
      • Gap1 antibody
      • GTPase activating protein antibody
      • HsCYK-4 antibody
      • ID GAP antibody
      • KIAA1478 antibody
      • Male germ cell RacGap antibody
      • MgcRacGAP antibody
      • Protein CYK4 homolg antibody
      • Protein CYK4 homolog antibody
      • Rac GTPase activating protein 1 antibody
      • Rac GTPase-activating protein 1 antibody
      • RACGAP 1 antibody
      • Racgap1 antibody
      • RGAP1_HUMAN antibody
      see all

    画像

    • Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      All lanes :

      Lane 1 : K562 cell lysate (in RIPA buffer)
      Lane 2 : Jurkat cell lysate (in RIPA buffer)

      Lysates/proteins at 35 µg per lane.

      Predicted band size: 71 kDa



      Primary incubation was 1 hour. Detected by chemiluminescence.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      ab2270 (1µg/ml) staining Human Testis by IHC-P. Microwaved antigen retrieval with Tris/EDTA buffer (pH9), HRP-staining.
    • Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      Anti-RACGAP1/MGCRACGAP antibody (ab2270) at 0.2 µg/ml + Human Testis lysate (RIPA buffer) at 30 µg

      Predicted band size: 71 kDa



      ab2270 staining (0.2µg/ml) of Human Testis lysate (RIPA buffer, 30µg total protein per lane). Primary incubated for 1 hour.  Detected by western blot using chemiluminescence.

      Primary incubated for 1 hour. Detected by western blot using chemiluminescence.

    • Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270)
      Anti-RACGAP1/MGCRACGAP antibody (ab2270) at 1 µg/ml + K562 cell lysate at 35 µg

      Developed using the ECL technique.

      Predicted band size: 71 kDa
      Observed band size: 75 kDa
      why is the actual band size different from the predicted?

    参考文献 (21)

    ab2270 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。

    ab2270 は 21 報の論文で使用されています。

    • Khalid M  et al. Gene Regulation by Antitumor miR-204-5p in Pancreatic Ductal Adenocarcinoma: The Clinical Significance of Direct RACGAP1 Regulation. Cancers (Basel) 11:N/A (2019). PubMed: 30866526
    • Pu J  et al. lncRNA MAGI2-AS3 Prevents the Development of HCC via Recruiting KDM1A and Promoting H3K4me2 Demethylation of the RACGAP1 Promoter. Mol Ther Nucleic Acids 18:351-362 (2019). PubMed: 31629962
    • Barbiero I  et al. CDKL5 localizes at the centrosome and midbody and is required for faithful cell division. Sci Rep 7:6228 (2017). PubMed: 28740074
    • Taulet N  et al. IFT proteins spatially control the geometry of cleavage furrow ingression and lumen positioning. Nat Commun 8:1928 (2017). PubMed: 29203870
    • Bornschein J  et al. Expression of aurora kinase A correlates with the Wnt-modulator RACGAP1 in gastric cancer. Cancer Med 5:516-26 (2016). IHC . PubMed: 26778597
    View all Publications for this product

    レビューと Q&A

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    1-6 of 6 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RACGAP1/MGCRACGAP antibody

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HT1080)
    Permeabilization
    Yes - 0.5% Triton X100
    Specification
    HT1080
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 21 2015

    Western blot abreview for Anti-RACGAP1/MGCRACGAP antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HT1080)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    20 µg
    Specification
    HT1080
    Blocking step
    Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 13 2015

    Western blot abreview for Anti-RACGAP1/MGCRACGAP antibody

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    20 µg
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jul 04 2013

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RACGAP1 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Methanol
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 27°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 16 2008

    Question

    I used Lysis buffer consisting of the following components: 1% Triton X-100, 0.5% NP-40, 150mM NaCl, 10mM Tris-HCL (pH 7.4), 1x protease inhibitor, 1 mM EDTA to lyse the cell pellets (liver cell lines harvested using 0.25% trypsin with EDTA) at 4oC for 40 to 60 min. Then the lysate were spun for 13K rpm at 4oC for 10 min. The supernatant was then transfered to a new tube and 1 ul was used for quantitation using Bio-rad Protein assay. I mix the protein( 40 ug each sample) with 5x sample buffer (with final concentration containing 0.3M Tris-HCL pH 6.8, 10% SDS, 30% Glycerol, 10% Mercaptoethanol, ~4% Bromophenol Blue) and ddH2 O to make up the final volume of 50 ul so as to give a final 1x sample buffer. The mixture were boiled (100oC) for 5 min and quick spun before loading onto SDS-PAGE gels (10% resolving and 4% stacking gels). After transfering onto Nylon membrane, the membrane was blocked using skim milk powder for about 40 to 60 min before being probed by 0.4ug/ml RACGAP1 antibody (ab2270) at 4oC for overnight. The next day , the blot was washed with 3 times 1xPBS containing 0.05% Tween-20 before being probed by secondary Ab (1:10k diluted rabbit anti-goat HRP conjugated Ab). The wash was repeated for 3 times before staining with 1: 1 mix of Immobilon Western HRP-substrate peroxide and HRP-substrate Luminol (Millipore) for 5 min at room temp. May I know the detail protocol on how to prepare the lysate (HumanTestis lysate) in the ab2270 datasheet and is it very different from what I performed (e.g: lysis, gel condition etc). I wonder the extra band (lower MW - between 38.3 kDa and 55.9 kDa) is the phophosrylated form of this protein?? Hear from you urgently.

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    Answer

    Thank you for your e-mail. The lysate used was Human Testis lysate lysed in RIPA buffer, the recipe for that buffer is as detailed below: 50mM Tris HCl pH 8 150 mM NaCl 1% NP-40 0.5% sodium Deoxycholate 0.1% SDS protease inhibitor cocktail (Roche or Sigma) I do not think the lower band can be the phosphorylated protein as phosphorylated proteins have relatively the same MW as non phosphorylated (if not they are a little higher but this is often not detectable by western blotting) and I think the lower band is more likely to be a degradation product (if your protease inhibitors are not efficient enough (not enough or too old)) or a fragment from post translational cleavage. I would recommend to run the positive control to see if you observe that band, and also to run a no primary control and to try blocking with BSA 5% 1 hour in case it is the blocking agent creating this problem. If you can also try overnight blocking in milk this may also be a good modification to try (if you have plenty of samples). Please let me know how you get on with those modifications and do not hesitate to contact me if you need further assistance,

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    Question

    Dear Abcam, I have a customer that is not getting a signal in the above antibody on HeLa cell extract. Do you know of any problems or any tips for this antibody. Thank you,

    Read More
    Answer

    Thank you for your enquiry. This antibody has been tested on Human Testis lysate, therefore we would strongly suggest using this type of lysate as positive control along with the samples. Should your customer still have difficulty with this product, then please provide some more details of his assay.

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