Anti-CD3 antibody (ab828)

Here is a link to our protocol for sodium azide removal

https://www.abcam.com/index.html?pageconfig=resource&rid=13869

Thank you for contacting Abcam.

We have not tested to see if ab828 would react with mouse cells. There is a chance that it would work but we cannot guarantee it. We do have a number of other CD3 antibodies in our catalogue that we know work with mouse samples, these include:

https://www.abcam.com/CD3-antibody-ab5690.html
https://www.abcam.com/CD3-antibody-ab16044.html



If however you would like to use ab828, then you may be interested in our 100% Abreview Testing Discount program:

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount



If there is anything else I can help you with, or if you would like to take advantage of our testing discount program, then please let me know.

Thanks for your call today and for your patience while I have looked into your enquiry. There is generally some issue with high background when using an antibody on homologous tissue, and several factors contribute to the problem: 1) The secondary antibody reacts with endogenous tissue IgG's (especially in tissue such as spleen that produces IgG's). The best way to reduce this is by blocking with an unconjugated anti-rabbit IgG F'ab fragment such as ab6824- https://www.abcam.com/Rabbit-IgG-secondary-antibody-H-L-Fab-Fragment-ab6824.html 2) The primary and secondary antibodies bind non-specifically to endogenous tissue Fc receptors (also especially in tissue like spleen). The Fc receptors should be blocked by the serum block that you are already using, but make sure to use at least 10% porcine serum and incubate the tissue in serum overnight at 4C to ensure that the receptors are blocked. 3) If using a biotinylated secondary antibody, the streptavidin-HRP may react with endogenous tissue biotin, so make sure to do an avidin/biotin block (this is probably included in your IHC kit if this step is necessary). Also make sure to do a peroxidase block, but you are probably already doing this as well. I think the most important step is to block the endogenous IgG with the anti-rabbit F'ab fragment. You also have the option of eliminating the need for a secondary antibody by directly conjugating the primary antibody to HRP. We have an EasyLink conjugation kit that allows you to do so- https://www.abcam.com/EasyLink-HRP-Conjugation-Kit-1-x-100µg-HRP-ab102890.html I hope this information will be helpful in resolving this issue, but if the problem persists please let me know and I will be happy to help further.