ELISA Question - Finding the concentration of Ab from cell culture (Feb/12/2007 )
HI all,
I did ELISA to find out the concentration of Ab in the cell culture supernatant. Supposing for some reason, I repeat the ELISA after a day or two, should I get the same data? Well, I don't. Can anyone throw some light on this?
Thanks.
May be
1. variation of aliquot
2. variation of dilution
3. variation of titration
The results done on different days should not have a hugh difference....should be around 10-15% difference....if lucky 5% or not so lucky 20%.
Hope this help.
1. variation of aliquot
2. variation of dilution
3. variation of titration
The results done on different days should not have a hugh difference....should be around 10-15% difference....if lucky 5% or not so lucky 20%.
Hope this help.
Hi Thank for your response. But generally do every one get within the above range of difference? If yes, then which one to consider provided the standards are perfect in both the experiments? Do we take an average? Thank you.
Hi!
If you want to avoid big deviation in results for a different days you first of all should keep your super with Mabs in the same conditions ( I mean aliquote your samples and freeze better -80 ( but not for a long peiod of time) And so must be done with antigen ( aliquote and freeze)
Today I don't recommend you to do any serious experiments ! Enjoy Saint Valentine's day today !
Also, do your incubations in a temp controlled incubator and watch your pipetting.