Coating ELISA Plate.. Tips please - (Mar/28/2008 )

Hello. I'm new to this ELISA work and I really need some advice and guidance on what is the best way to coat different amount of recombinant protein onto ELISA plate. I wish to coat my plate with different amount of protein starting with 0ng, 25ng, 50ng... to 200ng. But I need to know what is the best way to do this. Shall i do a serial dilution starting from 200ng, and dilute it or shall I prepare each amount of protein separately then coat it into the plate. which one is better?

Secondly, can anyone suggest any good website / books / papers on ELISA.

Thank you

Considering the very low weight of the substance, and economic reasons, it would be better to do a serial dilution. there would be less errors introduced by this approach.

As for the book, "current protocols in immunology" can be a good start.

good luck

I agree serial dilution is best. Use multiwellpipettor, wash wells vigorously, incorporate sucrose into your final blocking step (this will allow you to dry your plates and store at 4C. Coat at just above the pI of the protein.

Good luck.

Hi sqt4boston,
I read your reply saying to use sucrose so that the plates can be dried and stored at 4C

I need to do such process for ELISA
can you please give me the reference for that protocol
Leelaram