Blank controls in ELISA? - (Nov/01/2007 )

Hello,
jsut brifly, what should go in the 'blank' row on an ELISA plate? Is it all the reagents except the antigen?

So, i would add primary antibody, block, NO antigen, Secondary antibody and substrate??

Thanks!

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yes, no antigen, just culture medium or the buffer your standard/sample is in/diluted in. This should give your background reading.

Ceri

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That's strange,, In my lab as a control we use antigen WITHOUT primary antibody,,,
Which one is correct??

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the blank will depend on what you are coating on the plate; i'll try to explain better:
for Ab ELISA you'll coat with antigen and you blank is no primary antibody (ie the buffer where your antibody is, without the antibody)
for cytokine ELISA you coat with primary antibody, so blank is no antigen (ie: the buffer or media your sample is in).

hope this makes sense and helps.

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aah i see i see,,
yes that definitely helps! thanks.
i'm doing Ab ELISA so nothing wrong there

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Great, thanks everyone!

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