Question (45205) | Goat F(ab')2 Anti-Mouse IgG - (Fab')2, pre-adsorbed (ab98754)

Go to datasheet (ab98754)

Question

I have a question aboutab8978.
I found that it is mouse monoclonal and was tested in mouse sample on the data sheet.
I thinks that it is theoretically impossible because this antibody have to use anti-mouse IgGas secondary antibody.
So it can show high backgroundby non-specific bindingto endogenous mouse IgG especially in case ofIHC.
I would like to know how it is possible.
Please inform me.

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/Goat-Fab2-Mouse-IgG-Fab2-pre-adsorbed-ab98754.html (or use the following: https://www.abcam.com/Goat-Fab2-Mouse-IgG-Fab2-pre-adsorbed-ab98754.html).

https://www.abcam.com/Goat-Fab-Mouse-IgG-HL-ab6668.html (or use the following: https://www.abcam.com/Goat-Fab-Mouse-IgG-HL-ab6668.html).



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/Mouse-on-Mouse-Polymer-IHC-Kit-ab127055.html (or use the following: https://www.abcam.com/Mouse-on-Mouse-Polymer-IHC-Kit-ab127055.html).


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.

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