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

Submit a review Q&A (2)

Overview

  • Product name

    Anti-Shiga Toxin A subunit antibody [1C6A6G9]

  • Description

    Mouse monoclonal [1C6A6G9] to Shiga Toxin A subunit

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, ELISAmore details

  • Species reactivity


    Predicted to work with: Escherichia coli

  • Immunogen

    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

Properties

  • Form

    Liquid

  • Storage instructions

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

  • Storage buffer

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

  • Purity

    Affinity purified

  • Clonality

    Monoclonal

  • Clone number

    1C6A6G9

  • Isotype

    IgG1

Applications

Our Abpromise guarantee covers the use of ab101839 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000. Predicted molecular weight: 36 kDa.
ELISA Use at an assay dependent dilution.

Target

  • Relevance

    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).

  • Database links

    • Alternative names

      • Stx2 A subunit antibody

    References

    ab101839 has not yet been referenced specifically in any publications.

    Publishing research using ab101839? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    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.

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    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.

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    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

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    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.

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