Goat F(ab')2 Anti-Mouse IgG - (Fab')2, pre-adsorbed (ab98754)

Overview

  • Product name

    Goat F(ab')2 Anti-Mouse IgG - (Fab')2, pre-adsorbed
    See all IgG secondary antibodies
  • Host species

    Goat
  • Target species

    Mouse
  • Specificity

    Antiserum has been cross adsorbed against human and rat immunosorbents.
  • Tested applications

    Suitable for: ICC, IHC-P, ELISA, WBmore details
  • Minimal
    cross-reactivity


    Human, Rat more details

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

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

    Immunogen affinity purified
  • Purification notes

    This antibody was isolated by affinity chromatography using antigen coupled to agarose beads. F(ab')2 fragment were generated using a pepsin digestion. Fc fragments and whole IgG molecules have been removed.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab98754 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 Use at an assay dependent dilution.
IHC-P 1/200 - 1/2000.
ELISA 1/100 - 1/30000. Coating: Use 1/100-1/500 dilution; Primary: Use 1/1,000-1/30,000 dilution.
WB 1/1000 - 1/30000. Colorimetric: Use 1/1,000-1/10,000 dilution; Chemiluminescent: Use 1/1,000-1/30,000 dilution.

References

ab98754 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your patience.

I confirmed with the lab and am happy to forward their confirmation that ab98754 will detect both the (Fab) and the (Fab’)2 fragments of mouse IgG.

Please note that ab98754 itself is also a (Fab')2 fragment and therefore does not have an Fc part.If this is not suitable for the experiments that are planned, I am currently waiting on more information on ab139582. If the lab confirms that this antibody is also binding to both (Fab') and (Fab')2, I will let you know.

https://www.abcam.com/index.html?datasheet=139582 (or use the following: https://www.abcam.com/index.html?datasheet=139582).

I hope this information is helpful and wish you good luck with your experiments.

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Answer

Thank you for this information.

I have noted that this primary/secondary antibody combination has been used and reviewed by another customer. Their Abreview includes a detailed protocol which you may wish to review. This review is at:https://www.abcam.com/index.html?datasheet=33483&tab=abreviews&intabreviewid=14724

It may be in your case that much of the background is caused by secondary antibody binding to
endogenous mouse IgG in the tissue being stained, and to Fc receptors on B cells, plasma cells and macrophages. As such it may be valuable to use some of the staining procedures for mouse on mouse staining. Blocking endogenous IgG may be done by incubating the sections with an unconjugated Fab fragment likeGoat F (ab')2 polyclonal Secondary Antibody to Mouse IgG - (Fab')2, pre-adsorbed (ab98754),after the normal blocking step.More details may be found at this link:
https://www.abcam.com/index.html?pageconfig=resource&rid=11465

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Answer

Thank you for contacting us.

Indeed, a mouse-on mouse staining can be quite tricky; however, it is not impossible. The background will depend on the fixation method ( perfusion fixed tissue vs. normally fixed tissue), on the tissue itself ( eg. kidneys vs. bone) and the experimental settings.


One way to make sure to decrease or to completelyerase the background is to block the endogenous mouse IgG with a non- conjugated secondary antibody before you incubate with your target antibody. I would suggest to use a Fab or F(ab)2 fragment antibody in this case as they will bound in a higher density than whole IgG antibodies.

You can find the aforementioned antibodies here:


https://www.abcam.com/index.html?datasheet=98754 (or use the following: https://www.abcam.com/index.html?datasheet=98754).

https://www.abcam.com/index.html?datasheet=6668 (or use the following: https://www.abcam.com/index.html?datasheet=6668).



Another way is to use our hassle- free Mouse on Mouse (M.O.M) Polymer IHC Kit, which contains a special rodent block solution:

https://www.abcam.com/index.html?datasheet=127055 (or use the following: https://www.abcam.com/index.html?datasheet=127055).


In addition, we also received an Abreview (17 July 2009) from one of our customers who rated this antibody highly in IHC. You can contact the author via our website if you like.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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