I currently have rabbit primary AGO2 antibody; however, it does not give strong bands in mouse plasma samples and this antibody has never been tested on mouse samples before. I see AGO2 band, which weak with several unspecific bands.
I was wondering if anyone by experience knows of a good AGO2 antibody that works well in mouse plasma samples (without much unspecific binding/gives clear bands) and also is compatible for IP? (but not necessarily)
Thank you
Some tips for reducing non-specific binding and getting clear bands.
- I think you should increase your transfer time or increase the sample loading.
- Increase your incubation time in blocking buffer (5% w/v in Tris buffer saline Tween 20 (TBST) to get rid of unwanted bands if it appears. Prepare blocking solution fresh (just before use) and make sure the solution is properly mixed and is a uniform mixture with no clumps. Confirm that your blocking reagent is of proper quality (not expiry one).
- First fix your primary antibody dilution. If you are getting unwanted bands by adding primary antibody, then decrease dilution of primary antibody depending on the intensity of the band of interest. When intensity of your band of interest is appropriate, it is the proper primary antibody dilution. (fix this dilution when intensity of the band of interest is appropriate).
- Then try to standardize dilution of your secondary antibody. If you are still getting unwanted bands then increase the dilution of your secondary antibody until you don't get extra unwanted bands.Try to maintain temperature of 4°C during incubation with blocking buffer, primary and secondary antibody.
- Also confirm that your detection reagents (Reagent A and Reagent B of kits) are not expired and mix the two solutions only prior to use.
Not recommending from experience, but Wako's mAb anti-Mouse AGO2 has been around for a long time, and they say about it:
"This monoclonal antibody is not only used for western blot and immunohistochemistry, but also immunoprecipitation of mAGO2."
We also list a few other sources you may want to explore:
https://www.linscottsdirectory.com/antibodies?q=AGO2
you can filter for species reactivity and application and follow "more info" to each supplier's data page.
Dear Anar Alshanbayeva: I would propose Abcam Ab's, e.g.
Anti-Argonaute-2 antibody (ab32381) or Anti-Argonaute-2 antibody (ab186733) bothsuited for WB (see figures).
I would propose Abcam antibodies, e.g.:
Anti-Argonaute-2 antibody [EPR10411] (cat# ab186733).
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