Posted 17 July 2003 - 06:05 AM
I have a question regarding antibody stability. I have generated this antibody in chicken and affinity purified the antibody. It was working really well for a while and then it lost almost all activity. I tried purified more from serum and the purified antibody does not seem to be very stable. Immediately after purification, the antibody works well in immunostaining and western blotting. Soon after (even a week after), the activity starts to decrease and more background is detected. This seems very strange to me. I have purified other antibodies from goat and rabbit and they are very stable. (After the affinity column, I dialyzed the antibody with PBS and store it at either -80oC or 4oC with concentration at least 1 mg/ml) I am not sure what is causing the loss of activity (and such quick loss too!)
Anyone has any suggestion will be extremely welcome. Thank you.
Posted 21 August 2003 - 12:06 AM
2. don't freeze and thraw your Ab a lot. Try to aliquot your Ab into several eppendorfs. Use one at once.
Hope it could help you.
Posted 07 October 2003 - 04:59 AM
Don't add sodium azide to your Ab , which would inhibit your activity of HRP enzyme.