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Anti-Shiga Toxin A subunit 抗体 [1C6A6G9] (ab101839)

Reviews (1)Q&A (2)

Key features and details

  • Mouse monoclonal [1C6A6G9] to Shiga Toxin A subunit
  • Suitable for: ELISA, WB
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Shiga Toxin A subunit antibody [1C6A6G9]

  • 製品の詳細

    Mouse monoclonal [1C6A6G9] to Shiga Toxin A subunit

  • 由来種

    Mouse

  • アプリケーション

    適用あり: ELISA, WBmore details

  • 種交差性


    交差が予測される動物種: Escherichia coli

  • 免疫原

    Synthetic peptide (

    REFRQALSETAPVYT

    ) corresponding to amino acids 198-212 of Shiga Toxin A subunit.
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 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. Avoid freeze / thaw cycles.

  • バッファー

    pH: 7.20
    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • 精製度

    Affinity purified

  • ポリ/モノ

    モノクローナル

  • クローン名

    1C6A6G9

  • アイソタイプ

    IgG1

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ab101839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
ELISA
Use at an assay dependent concentration.
WB (1)
Use at an assay dependent concentration.
特記事項
ELISA
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.

ターゲット情報

  • 関連性

    Shiga toxins are a family of related toxins with two major groups, Stx1 and Stx2, whose genes are considered to be part of the genome of lambdoid prophages. The most common sources for Shiga toxin are the bacteria S. dysenteriae and the Shigatoxigenic group of Escherichia coli (STEC), which includes serotype O157:H7 and other enterohemorrhagic E. coli (EHEC).

  • 参照データベース

    • 別名

      • Stx2 A subunit antibody

    参考文献 (0)

    ab101839 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。

    ab101839 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

    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-Shiga Toxin A subunit antibody [1C6A6G9]

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Escherichia coli Cell lysate - whole cell (whole cell)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (15% gel)
    Loading amount
    5 µg
    Treatment
    0.2% L-arabinose for 4hrs
    Specification
    whole cell
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Bing Chang Yang

    Verified customer

    投稿 Oct 07 2020

    Question

    Are they using the purified protein or anEHEC O157:H7 lysate? If the later, I would suggest to increase the protein load up to 20 to 30ug per lane to offer the antibody enough target protein.
    - They used lysate and it's hard to get enough sample, he concentrated the protein and applied. Protein sample loaded 15ug/lane and he said he couldn't loading more than that because sampling was difficult.

    How was the transfer verified?
    -It was checked by ponceau staining.

    For a better saturation of the membrane, I would recommend a blocking buffer containing 3% BSA for 2 hours at room temperature. BSA can sometimes give better results than milk. If you are going to switch to BSA, please switch the incubation buffer as well.
    -This is not about non-specific band so the customer thinks it'snot necessary to try BSA blocking

    Could you please confirm the host species of the secondary antibody and the species it reacts against?
    -anti-mouse IgG(HRP)


    They checked DNA level byPCR and this WB is for last confirmation.

    Read More
    Answer

    Thank you for your reply.


    Indeed, usually the blocking step is connected with the occurrence of side bands. However, changing the blocking agent can significantly increase the signal strength as our lab found out. I have attached a PDF which shows what can happen when you use the same material and protocol except for the blocking agent.


    I hope this information is helpful to you.If the results do not get better, we will certainly replace the product.

    Read More

    Question

    LOT NUMBER gr37811-4 ORDER NUMBER 1013227 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE EHEC O157:H7 PRIMARY ANTIBODY Concentration or dilution: 1/250-1/1000 Diluent buffer: 2% skim milk Incubation time: o/n (˜18 hrs) Incubation temperature: 4℃ Buffer: 0.1% Tween-TBS Number of washes: 3 times for 10 min DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Positive control Stx toxin negative control no ANTIBODY STORAGE CONDITIONS -20℃ SAMPLE PREPARATION Lysis buffer: no Protease inhibitors: sigma, protease inhibitor cocktail Phosphatase inhibitors: no Reducing agent: 14.4 mM b-mecaptoethanol Boiling for ≥5 min? yes AMOUNT OF PROTEIN LOADED 1-5 ug per lane ELECTROPHORESIS/GEL CONDITIONS sds-page, 10% TRANSFER AND BLOCKING CONDITIONS Type of membrane: nitrocellulose Protein transfer verified: yes Blocking agent and concentration: 5% skim milk Blocking time: 2 hrs Blocking temperature: RT SECONDARY ANTIBODY Species: mouse Isotype: IgG Reacts against: anti-mouse Concentration or dilution 1/2500, 1/1000 Diluent buffer: 2% skim milk Incubation time: 4 hrs Incubation temperature: 37℃ Fluorochrome or enzyme conjugate: HRP Buffer: 0.1% Tween-TBS Number of washes: 3 times for 10 min 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? Concentrations and incubation time of primary antibody and secondary antibody. ADDITIONAL NOTES no band

    Read More
    Answer

    Thank you for taking time 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.

    Having reviewed this case, I would like to offer some suggestions to help optimize the results fromab101839. I would also appreciate if you can confirm some further details:

    Are they using the purified protein or anEHEC O157:H7 lysate? If the later, I would suggest to increase the protein load up to 20 to 30ug per lane to offer the antibody enough target protein.
    How was the transfer verified?
    For a better saturation of the membrane, I would recommend a blocking buffer containing 3% BSA for 2 hours at room temperature. BSA can sometimes give better results than milk. If you are going to switch to BSA, please switch the incubation buffer as well.
    Could you please confirm the host species of the secondary antibody and the species it reacts against?





    Should the suggestions not improve the results, please do let me know.

    In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

    I hope this information is helpful, and I thank you for your cooperation.

    Read More

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