Goat Anti-Mouse IgG H&L (Biotin) (ab6788)

Submit a review Q&A (7)References (37)

Overview

  • Product name

    Goat Anti-Mouse IgG H&L (Biotin)
    See all IgG secondary antibodies

  • Host species

    Goat

  • Target species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, ICC, Dot blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WBmore details

  • Immunogen

    mouse IgG whole molecule

  • Conjugation

    Biotin

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.878% Sodium chloride, 1% BSA, 0.424% Potassium phosphate
  • Concentration information loading...

  • Purity

    Affinity purified

  • Purification notes

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • General notes

    The conjugate is expected to have between 5 and 10 moles of biotin per mole of IgG.

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6788 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
ICC/IF 1/1000 - 1/5000.
ICC Use at an assay dependent dilution. PubMed: 17980964
Dot blot Use at an assay dependent concentration.
ELISA 1/350000.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Immunomicroscopy Use at an assay dependent concentration.
WB 1/2000 - 1/20000.

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Mouse IgG H&L (Biotin) (ab6788)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Mouse IgG H&L (Biotin) (ab6788)

    IHC image of Histone H1 staining in a section of formalin-fixed paraffin-embedded human normal colon*.

    The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6) for 30 mins. The section was then incubated with ab11080, 1/1,000 dilution, for 15 mins at room temperature. A goat anti-mouse biotinylated secondary antibody (ab6788, 1/1000 dilution), was used to detect the primary, and visualized using an HRP conjugated ABC system. Streptavidin HRP was used, ab7403 at a 1/10,000 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10 min at room temperature. The section was then counterstained with hematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Mouse IgG H&L (Biotin) (ab6788)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Mouse IgG H&L (Biotin) (ab6788)

    ab6788 was used at dilution 1/500 with the primary antibody ab2866 in IHC-P. See the review on ab2866.

  • ELISA - Goat Anti-Mouse IgG H&L (Biotin) (ab6788)
    ELISA - Goat Anti-Mouse IgG H&L (Biotin) (ab6788)

    ELISA - Goat Anti-Mouse IgG H&L (Biotin) (ab6788)

    ELISA results of purified ab6788 tested against purified Mouse IgG. Each well was coated in duplicate with 1.0 µg of Mouse IgG. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log 10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gelatin as blocking buffer, Streptavidin Peroxidase Conjugated and TMB substrate.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References

Publishing research using ab6788? Please let us know so that we can cite the reference in this datasheet.

ab6788 has been referenced in 37 publications.

  • Hughes LJ  et al. Magnitude and diversity of immune response to vaccinia virus is dependent on route of administration. Virology 544:55-63 (2020). PubMed: 32174514
  • Tang Z  et al. Antitumor Drug Combretastatin-A4 Phosphate Aggravates the Symptoms of Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice. Front Pharmacol 11:339 (2020). PubMed: 32265711
  • Yao Y  et al. Knockdown of long noncoding RNA Malat1 aggravates hypoxia-induced cardiomyocyte injury by targeting miR-217. Adv Clin Exp Med 28:719-728 (2019). PubMed: 30843674
  • Foyt DA  et al. Hypoxia impacts human MSC response to substrate stiffness during chondrogenic differentiation. Acta Biomater 89:73-83 (2019). PubMed: 30844569
  • Yu B  et al. LPS-induced upregulation of the TLR4 signaling pathway inhibits osteogenic differentiation of human periodontal ligament stem cells under inflammatory conditions. Int J Mol Med 43:2341-2351 (2019). PubMed: 31017254
View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Question

The secondary antibody ab6788 gives no signal. We have compared it to an effective anti-mouse IgG secondary, also biotinylated.

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Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question

Hi Abcam,

Do you mind helping with this enquiry below?

We have found another couple of secondary antibodies on your website that appear identical to AB6788 & AB6720 except for a different concentration.

They are: ab97049 Goat anti-rabbit IgG H&L (biotin)

and ab 97021 Goat polyclonal Secondary antibody to mouse IgG- H&L (Biotin)

What we are hoping to do is to enhance a western blot signal by adding a streptavidin step prior to applying a biotinylated secondary antibody. (This has been successfully done by colleagues in the past).

Would it be possible to get some advice as to which ones would be more suitable for our application?

Thanks for your help :D

Kind regards,

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Answer

Thank you for contacting us and you interest in our products.

There are a few differences between the two pairs of antibodies you have identified. A few of these include the purification used (affitiny vs IgG fraction) as well as the ratio of conjugation. The ab6788 and ab6720 is estimated to have a conjugation ratio of 10-20 per IgG molecule whilst ab97049 and ab97021 have a ratio of 1:4.

I am not familiar with the amplification technique you intend to use and cannot therefore anticipate how these differences may alter the results.

If you could share a more detailed description of the procedure I may be able to advise in more detail.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

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Question

I am interested in purchasing a secondary antibody for use in immunohistochemistry that can work with the following primary antibodies from your company (ab24590, ab7817, ab955, and ab89064). I have searched the catalog and found ab6728 and ab6788 as possible candidates for this purpose. Could you kindly suggest on which one would work better with those primary antibodies for immunohistochemistry (IHC-P)? Thank you,

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Answer

Thank you for contacting Abcam.

I feel that both products are suitable for IHC-P. Both have many specific references and in-depth scientific support data right on the datasheet. They differ mainly in the species in which they were raised and in the tags used for detection. Ab6728 is an HRP conjugate whereas Ab6788 is Biotin labeled. I believe that you should choose the secondary which has an associated detection system that you are most familiar with in case you should run into any staining issues.

I hope this information is helpful. Please let me know if you have any questions.

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Question

Dear Sir/madam, Thank you very much for your kind attention. Secondary antibody that I used it is: Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Biotin) (ab6788) I have used the same secondary antibody with another mouse monoclonal Antibody and it works very well. Thank you very much again with my best regards.

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Answer

Thank you very much again for your reply. I appreciate the time you have spent in the laboratory and understand your concerns. As it appears to me your biotin-labeled secondary and the appropriate detection system are indeed working and it is regrettable the results have not been successful with the Progerin antibody yet. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial of ab66587. I apologise for the inconvenience and am pleased to offer you a free of charge replacement (with either a new vial or any other primary antibody of your choice) or credit note in compensation. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question

Dear Sir/madam, Thank you very much for your kind attention. These are the answers of all question: 1- No results, I mean that I obtain no bands at all. 2- a-The protein was denatured first. b- 5% milk c- 1:100 dilution d- Anti-mouse IgGBiotin secondary Abcam ab6788 Goat Thank you very much again with my best regards.

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Answer

Thank you very much for confirming these details. Before proceeding with this case, I would appreciate if you could confirm which detection system was used with the biotin-labeled secondary antibody ab6788: Have you used a HRP-conjugated anti-biotin antibody or HRP-conjugated streptavidin, and is this working for other primary antibodies in your lab? I look forward to your reply.

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Question

Hi, Could you please tell me the molecular weight of the antibody ab6788 in either daltons or grams (IgG is supposed to be 150,000 daltons in humans. But this one is from rabbit and has other modifications to it.) Also, your online information sheet says Biotin/protein ratio as 10 - 20. Does this mean there are 10 - 20 biotin molecules per Ab? Thank you

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Answer

Thank you for your enquiry. Unfortunately, the molecular weight of this product is not specifically measured, but the molecular weight of non reduced IgG is approximately 150-160 kDa. The molecular weight of biotin is ~250 Daltons. Yes, there are 10-20 molecules of Biotin conjugated to the IgG. The ratio of 10-20 is based on the combination ratio and typical results from our standard conjugation protocol. Function and purity of intact conjugate is confirmed by Immuno-electrophoresis (with anti biotin) and ELISA against mouse IgG using a HRP conjugated streptavidin as a secondary reagent. I hope　this information is helpful. If there is anything else that I can help you with, please do not hesitate to contact me.

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Question

Thank you very much. Could you please let me know whether I can use PBS buffer as antibody diluent( my second query of ref:914295)

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Answer

Thank you for getting back to me. Phosphate buffered saline can be used as an antibody diluent as an alternative to TBS for western blot analysis. However, with regards storage I would recommend that the antibodies are alituotted and frozen at -20oC upon arrival without prior dilution. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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