Overview

  • Product name

    Sheep Anti-Mouse IgG H&L (HRP)
    See all IgG secondary antibodies
  • Host species

    Sheep
  • Target species

    Mouse
  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with mouse IgG and with light chains common to other mouse immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. This antibody may cross react with IgG from other species.
  • Tested applications

    Suitable for: ICC, ELISA, IHC-P, WBmore details
  • Conjugation

    HRP

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

    Preservative: 0.1% Proclin
    Constituents: 0.2% BSA, PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to HRP.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab102458 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.
ELISA 1/10000 - 1/100000. (Primary)
IHC-P 1/200 - 1/5000.
WB 1/10000 - 1/50000. with a chemoluminescence detection method. If using a colorimetric method, dilute 1/5000 - 1/30000.

References

ab102458 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

I have attached our dot blot protocol. However, on second thought, it may not be applicable to a blot of just the secondary, because I am not sure how well the HRP will handle the drying step that is at at the start of the attached protocol, after applying a protein sample to the nitrocellulose. You need the HRP to be active and drying the secondary is not equivalent to what happens when you apply the secondary in a standard WB procedure. (It never drys out in that procedure. On the other hand, the Pierce dot blot of HRP worked. I expect drying is necessary in order to keep it from washing of the blot).

Whatever you decide to do, just let me know whether you would like to try a different secondary or a credit or refund, and I will arrange that for you.

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Answer

Thank you for bringing this to our attention. It could very well be that the HRP is the issue, or the antibody:antibody affinity. I am assuming you are using the same primary with both secondaries (the more expensive one from another company and ours).

Without the data from the other secondary, I would say that the fading of chemiluminescent signal is expected. The depletion of substrate is possible, and diluting the antibody might address that, but I would expect that depletion would occur with the other secondary too, if the intensity of the initial signal was comparable to the ab102458 signal.
As I mentioned, if you are interested in trying one of the secondaries in our catalogue that have more sales, I will be happy to send one as a free replacement, for instance ab6789.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=6789).

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