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Anti-SH2D1B 抗体 (ab67417)

Submit a review Q&A (1)
Western blot - Anti-SH2D1B antibody (ab67417)

    Key features and details

    • Mouse polyclonal to SH2D1B
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG

    製品の概要

    • 製品名

      Anti-SH2D1B antibody
      SH2D1B 一次抗体 製品一覧

    • 製品の詳細

      Mouse polyclonal to SH2D1B

    • 由来種

      Mouse

    • 特異性

      This antibody is reactive against mammalian transfected lysate.

    • アプリケーション

      適用あり: WBmore details

    • 種交差性

      交差種: Human

    • 免疫原

      Recombinant full length protein within Human SH2D1B. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements.

    • ポジティブ･コントロール

      • SH2D1B transfected 293T cell lysate

    • 特記事項

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

    • バッファー

      pH: 7.4
      Constituent: PBS
    • Concentration information loading...

    • 精製度

      Protein A purified

    • 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

    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ab67417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
    1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
    特記事項
    WB
    1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).

    ターゲット情報

    • 関連性

      SH2D1B binds phosphotyrosines through its free SRC homology-2 domain. It regulates signal transduction through receptors expressed on the surface of antigen-presenting cells.

    • 参照データベース

      • 別名

        • EAT 2 antibody
        • EAT2 antibody
        • EWS/FLI1 activated transcript 2 antibody
        • SH2 domain containing 1B antibody
        • SH2 domain containing molecule EAT2 antibody
        • SH2 domain containing protein 1B antibody
        see all

      画像

      • Western blot - Anti-SH2D1B antibody (ab67417)
        Western blot - Anti-SH2D1B antibody (ab67417)
        All lanes : Anti-SH2D1B antibody (ab67417) at 1/500 dilution

        Lane 1 : SH2D1B transfected 293T cell lysate
        Lane 2 : Non-transfected 293T cell lysate

        Lysates/proteins at 25 µg/ml per lane.

        Secondary
        All lanes : Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

        Predicted band size: 15 kDa
        Observed band size: 15 kDa

      参考文献 (0)

      ab67417 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。

      ab67417 は論文での使用が確認できていません。

      レビューと Q&A

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      Question

      LOT NUMBER XXXXX ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Multiple bands or non-specific bands. SAMPLE Cell lysates of: A20 cell line (mouse), Devernelle cell line (human), mouse lymphocytes. PRIMARY ANTIBODY Anti-SH2D1B antibody (ab95270) abcam. Diluted 1/250 in PBS-Tween 0,1%. (Previously tritated) Incubated during 2 hours (when we tried 1 hour we observed NO bands)at room temperature in agitation. Washed three times during 10 minutes with PBS-Tween 0,1% at room temperature. DETECTION METHOD ECLPlus. POSITIVE AND NEGATIVE CONTROLS USED Positive: Jurkat cell lysate Negative: mouse serum lysate ANTIBODY STORAGE CONDITIONS 4°C, NOT sub-aliquoted. SAMPLE PREPARATION Cells are lysed with the following lysing buffer heated at 100°C prior mixing with the cells (we use a ratio of 8 million cells for 300 uL of lysing buffer): -SDS 10% -Tris-Cl 1M pH 7,4 -PMSF 100 mM -Na3VO4 0,1 M -Calbiochem protease inhibitor cocktail AMOUNT OF PROTEIN LOADED 20 ug and 40 ug (in both cases we obtain the same multiple bands) ELECTROPHORESIS/GEL CONDITIONS 12% polyacrilamide gel. Run at 50V during 30 min, then 100V for 1 hour. Using Electrophoresis buffer (Tris 25mM, Glycine 0,2M, SDS 0,1%) TRANSFER AND BLOCKING CONDITIONS Transferred during 2 hours at 4°C, using Transfer Buffer (Tris 25mM, Glycine 0,2M, Methanol 20%. Bloking agent: PBS-Tween 0,1% + 3% BSA, blocked over night at 4°C. SECONDARY ANTIBODY DakoCytomation Polyclonal Goat Anti-Mouse Immunoglobulins/HRP Code No./ Code/ Code-Nr. P 0447 Diluted 1/5000 with PBS-Tween 0,1%. Incubated for 1 hour at room temperature in agitation and in darkness. Wash step: washed three times for 10 minutes at room temperature with PBS-Tween 0,1%. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 30 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

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      Answer

      Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. Reviewing this case, I would like to offer some suggestions to help optimise the results from ab67417. I would also appreciate if you could send me an image of any of the failed blots, in order to better understand why this antibody did not perform as stated on the datasheet. - Could you please confirm the order number? It will help us to track the -Attending your protocol, both Mouse and Human samples have been used, however, this antibody has only been characterised using human cell lines, and we don’t have any evidence of its suitability for Mouse samples. -I would recommend using less concentrated antibody solutions, as 1/500 and 1/1000, and let the antibody incubate at 4 C overnight. -For the sample preparation, we suggest using RIPA buffer, which has a higher denaturing level. The recipe to prepare it can be found under the link: https://www.abcam.com/index.html?pageconfig=resource&rid=11414 It is also important to use stronger reducing conditions, supplementing the lysis buffer with DTT or mercaptoethanol, and heating the sample at 95C for 5 minutes. -It would be interesting to try different blocking conditions, as, 5% of non fat milk in TBST, for 1-2 hours at RT. -Regarding the antibody storing conditions, if no cycles of freezing/thawing are done, the antibodies are viable for up to one year at 4C, so in this particular case, if the antibody was stored for 3 months before being used, there shouldn’t be any degradation issue. I hope the above recommendations may help you, if you still experience problems please do not hesitate to contact me with an image of the blot if possible. I would also appreciate if you could also provide details of your order (purchase order number, date of purchase, shipping address/purchasing agent information) in order to dismiss any shipping issue, as well as to monitor the concrete lot complaints. Please do not hesitate to contact us if you need any more advice or information.

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