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I use your protocol for BrdU staining. Please see attached file. I used your antibody, ab2284 on mice intestine tissue last November, I got beautiful staining. This month, we are doing another batch of mice and following exactly protocol as November’s. I did three run of staining to exclude any mistake I might have and still got no stain at all. Finally, I thought it was DAB problem and emailed you last week. After receiving new DAB, I repeat BrdU stain with a fresh aliquated antibody. Then I got faint stain - comparing with last November’s staining. At that time, I thought it was Streaptavidin problem and emailed you for that. Since I need result quickly, I run another run with my real slides on Monday with the antibody I thawed last Thursday (got result on Friday) and got no stain at all even with new DAB. This antibody was aliquated and stored at -200C since we received last September. This particular tube was thawed last Thursday and used only once. It was put back to -200C right after usage. This Monday was the second time to thaw and to use, and I did not get any staining, the only explanation is that the antibody got degraded.
I hope you agree with me or give me some suggestion. I also hope you could send me new antibody to see what I get.
Asked on Mar 29 2012
Thank you for your reply and forsending the additional information.
It does sound like the antibody was damaged during storage, so I am happy to send a replacement vial for you. This will be coming free of charge on the order ***, which should arrive tomorrow. Please note that we recommend storing this antibody in working aliquots, so that each aliquot is not frozen and thawed more than one time.
I apologize for the trouble with these products, but please keep me updated about how this replacement antibody works in your staining. Let me know if you have any questions or if there is anything else that we can do for you.
Answered on Mar 29 2012