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HRP Anti-Phosphoserine 抗体 (ab9334)

Submit a review Q&A (7)References (12)
Western blot - HRP Anti-Phosphoserine antibody (ab9334)

    Key features and details

    • HRP Rabbit polyclonal to Phosphoserine
    • Suitable for: WB, ELISA
    • Reacts with: Species independent
    • Conjugation: HRP
    • Isotype: IgG

    製品の概要

    • 製品名

      HRP Anti-Phosphoserine antibody
      Phosphoserine 一次抗体 製品一覧

    • 製品の詳細

      HRP Rabbit polyclonal to Phosphoserine

    • 由来種

      Rabbit

    • 標識

      HRP

    • 特異性

      Specifically recognizes both free-phosphoserine, serine-phosphorylated peptide and proteins. Does not cross-react with ATP, phosphotyrosine, peptidyl phosphothreonine and serine. Has slight cross-reactivity with free phosphothreonine. Readily reacts to known phosphoproteins such as phosvitin and alpha casein.

    • アプリケーション

      適用あり: WB, ELISAmore details

    • 種交差性

      交差種: Species independent

    • 免疫原

      Chemical/ Small Molecule corresponding to Phosphoserine conjugated to keyhole limpet haemocyanin. BSA and KLH-phosphoserine conjugates.

    • ポジティブ･コントロール

      • Use Mouse brain extract for Western Blotting or synthetic phosphopeptide or phosvitin for ELISA.

    • 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

    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ab9334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
    Use a concentration of 2 µg/ml. To block use 3%BSA with 0.1% gelatin (do not use milk). We recommend that the antibody solution should contain 0.5% BSA to prevent non-specific binding.
    ELISA
    Use a concentration of 1 µg/ml.
    特記事項
    WB
    Use a concentration of 2 µg/ml. To block use 3%BSA with 0.1% gelatin (do not use milk). We recommend that the antibody solution should contain 0.5% BSA to prevent non-specific binding.
    ELISA
    Use a concentration of 1 µg/ml.

    ターゲット情報

    • 関連性

      Changes in the serine/threonine phosphorylation state of a protein in response to various external stimuli can have profound effects on cellular signal transduction, apoptosis and carcinogenesis. The reagents, including phosphorylated protein/peptides, antibodies against the phosphospecific amino acid, are important tools to explore the activation of serine, threonine or tyrosine containing proteins. An aberrant protein phosphorylation is a hallmark of human disease, and the enzymes, particularly protein kinases, which control protein phosphorylation are recognized as a major new drug target family.

    • 別名

      • phospho-Ser antibody
      • pS antibody
      • pSER antibody

    画像

    • Western blot - HRP Anti-Phosphoserine antibody (ab9334)
      Western blot - HRP Anti-Phosphoserine antibody (ab9334)
      Western blotting of mouse brain extract with ab9334 at a 1/200 dilution on a 12% SDS gel. 50 µg protein loaded in each lane. Antibody incubated for 2 hours incubation at room temperature.

    参考文献 (12)

    ab9334 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。

    ab9334 は 12 報の論文で使用されています。

    • Liu X  et al. Pre-miRNA Hsa-Let-7a-2: a Novel Intracellular Partner of Angiotensin II Type 2 Receptor Negatively Regulating its Signals. Int J Biol Sci 18:3237-3250 (2022). PubMed: 35637969
    • Qu L  et al. Plant casein kinases phosphorylate and destabilize a cyclin-dependent kinase inhibitor to promote cell division. Plant Physiol 187:917-930 (2021). PubMed: 34608955
    • Wilkes MC  et al. Diamond Blackfan anemia is mediated by hyperactive Nemo-like kinase. Nat Commun 11:3344 (2020). PubMed: 32620751
    • Martínez-Florensa M  et al. Serine residues in the LAT adaptor are essential for TCR-dependent signal transduction. J Leukoc Biol 89:63-73 (2011). WB . PubMed: 20940326
    • Zang ZJ  et al. Identification of PP2A as a novel interactor and regulator of TRIP-Br1. Cell Signal 21:34-42 (2009). WB . PubMed: 18940248
    View all Publications for this product

    レビューと Q&A

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    レビューを送る 質問を送る

    1-7 of 7 Abreviews or Q&A

    Question

    unser Telefongespräch wurde gerade eben von einem Stromausfall in der Uni Bielefeld unterbrochen.

    Deshalb melde ich mich jetzt per E-Mail (mein PC hat Notstromversorgung).

    Ich wollte Sie noch fragen ob Sie uns ein Anwendungsprotokoll für das Blotten mit dem ab9334 schicken können.

    Read More
    Answer



    Es gibt eigentlich kein besonderes Protokoll für diesen Antikörper. Die einzigen Abweichungen von unserem Etablierungsprotokoll sind der Vermerk bezüglich des Blockmittels ("To block use 3%BSA with 0.1% gelatin (do not use milk). We recommend that the antibody solution should contain 0.5% BSA to prevent non-specific binding.") und die Inkubation für zwei Stunden bei RT. Unser Standardt-Protokoll finden Sie als Anhang an diese Email.

    Ich hoffe, dies hilft Ihnen trotzdem weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Question

    No signal. Any problems? Suggested a test with a secondary antibody to check if the abs or the conjugate may be the problem

    Read More
    Answer

    Vielen Dank für Ihren Anruf.

    Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten. Wie am Telefon besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 123456und solltemorgen bei Ihnen ankommen.

    Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Question

    No signal. Any problems? Suggested a test with a secondary antibody to check if the abs or the conjugate may be the problem

    Read More
    Answer

    Es tut mir leid zu hören, dass Sie Probleme mit diesem Antikörper haben.

    Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.




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    Question

    Our customer sent me additional data about your email.
    Check this please and he would like to receive replacement vial with different lot#.

    Best regards,

    Read More
    Answer

    Thank you very much for following up with this customer. I am sorry that the results were not improve with the suggestions.

    As requested, I have added a new free of charge unit of ab9334 of a different batch to your order xx

    I hope the results will be good with the new vial. Please do not hesitate to contact us again should you have any questions or concerns.

    Read More

    Question

    I'm seeing a black blot in WB using this antibody.

    Read More
    Answer

    Thank you for your inquiry.

    I am sorry to hear that you are experiencing difficulty with this antibody. Like we said on the phone, please try diluting the primary antibody more to 1:10,000 for 1 -2 hrs at RT, increasing the blocking to 1% BSA with the primary and trying regular ECL instead of the super signal.

    If these protocol modifications do not work to improve the signal, please let us know and we will look into replacing or refunding the product in accordance with our Abpromise. We look forward to hearing from you.

    Read More

    Question

    LOT NUMBER xx ORDER NUMBER xx DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE purified protein from sigma PRIMARY ANTIBODY Concentration or dilution 1/100 dilution (2.5ug/ml) Diluent buffer 0.5% BSA in PBST Incubation time overnight (16hrs) Incubation temperature: 4 degree C wash buffer PBST Number of washes 10min RT, 3 times DETECTION METHOD LAS4000 chemiluminescence POSITIVE AND NEGATIVE CONTROLS USED Positive control Beta-casein (Sigma, C6905) Negative control not used ANTIBODY STORAGE CONDITIONS -20 degree C SAMPLE PREPARATION Lysis buffer : not used (resuspension buffer : 20mM Tris-Cl pH 8.0) Protease inhibitors: not used Phosphatase inhibitors : not used Reducing agent DTT Boiling for 5 min? yes/no yes AMOUNT OF PROTEIN LOADED 1ug/lane ELECTROPHORESIS/GEL CONDITIONS 10% Tris-glycine TRANSFER AND BLOCKING CONDITIONS Type of membrane PVDF Protein transfer verified yes (with prestained marker) Blocking agent and concentration 3% BSA, 0.1% gelatin in PBST Blocking time 2hrs Blocking temperature RT SECONDARY ANTIBODY not used HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A NO PRIMARY CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? - ADDITIONAL NOTES -

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    Answer

    Thank you for taking time to liaise with the customer and to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

    I have reviewed this case and would like to confirm the following points with the customer:

    1.) In the image provided, the band of the casein protein seems to have a strange migration. Has the customer tested this casein protein on another gel with the same results?

    2.)As general suggestion, I can advise to keep the samples all the time on ice for detection of phosphoprotein. I do also recommend the use of phosphatase inhibitors. As I suppose however that the MS data and the gel have been performed with the same sample of casein, I do acknowledge that the casein protein has been phosphorylated and that therefore the positive control should provide a signal. See also our protocol for phosphoproteins. https://www.abcam.com/ps/pdf/protocols/Western%20blotting%20of%20phospho-proteins%20protocol.pdf

    In the event thatthis antibodyhas not worked on different blots detecting the phosphorylated casein, we would be pleased to provide a free of charge replacement orcredit note.

    I am looking forward to hear back from you.

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    Question

    I would like to use the Phosphoserine antibody HRP conjugate (ab9334)to recognize the phosphorylated serine in the following peptide: Biotin-ADVKFSFQCPG. Should this antibody work with this peptide or does it need specific surrounding amino acid? Thanks for your help.

    Read More
    Answer

    The phosphoserine antibodies were tested positive for the following phosphopeptides: RREILSRRPpSYR RRALpSLR TPRpSP

    Read More

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