Anti-PADI4 / PAD4 antibody (ab50247)

Thanks for your quick reply! I incubate membrane in PAD4 antibodies (1:1000) at 4 degree, overnight. The next day, wash with PBS-tween 5 min,X3 times, incubate with rb secondary Ab for 2h at room temperature, wash with PBST 5 min X 3, and then develop. I use this protocol for other antibodies and they work well. Thanks for passing my comments to the relevant team, and hopefully in the future I can get this Ab that's guaranteed to work with mice.

ANSWER:

Thank you for your reply.

There are a few variations that may work:

1 - Depending on the blocking agent you are using, you could try using 5% BSA, as this may help reduce the background levels that you are seeing.

2 - If you are seeing the band of interest, but also getting high background, then using a lower concentration of the antibody may be helpful in reducing background..

3 - Another, way to decrease background is to use ab126587, our http://www.abcam.com/10X-Blocking-Buffer-ab126587.html, but I would only recommend that if using the BSA does not prove to be successful.

Please let me know if there is anything else I can help you with.

I ordered PAD4 antibody (ab50247). It says it reacts with human, but predicted to react with mice. I tried in on mice tissue by Western Blot, but it doesn't work very well, and there're a lot of unrelated bands there. The antibodies we previously ordered from your company are very good, and I'm wondering whether you guys could produce PAD4 antibody that react with mice better. Thank you very much!
Best,

ANSWER:

Thank you for contacting Abcam.

If you could provide me with the protocol that you are using, I would be happy to see if there are any protocol suggestions I can make that may help resolve some of the issues we are having.

Also we have a number of other PADI4 / PAD4 antibodies in our catalogue that may work for mouse but we have not tested them, therefore you could take advantage of our testing discount program:

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



I will pass your comments about getting an antibody that we guarantee to work with mice to the relevant team.

If there is anything else I can help you with, please let me know.