Hey Everyone,
For the people with experience in biochemistry/ELISA.
what do you do in order to prepare human blood to see measure a protein through ELISA that is not highly concentrated in blood.
Do you get rid of albumin? if this is the case, which kit or protocol do you recommend?
thanks everyone for your tips and of course time.
Carolina
You may think about use of centricon tubes of required size... Suppose your sample is 40 KD. Then go for a 30 KD cut first... Get read of the effluent. Then resuspend the pellet and go for a 50 KD cut off spin... Take the effluent of 2nd spin and do your ELISA.
Thanks Diptiman
Yes, I have thought about using amicon filters, any oder suggestion?
The ultrafiltration looks like a good option, yet in commercial ELISAs many of the dilution buffers are loaded with albumin and also the coated plates are most often blocked with albumin as to prevent aspecific signals. If you are doing this in an experimental setup and not in routine (higher througput) analysis ypou might consider Western Blot to get rid of background occuring in the ELISA.
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