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ELISA Assay Forum ELISA Forum. Discuss and post questions including optimization, troubleshooting, protocols and theory of ELISA assays using monoclonal, and polyclonal antibodies or peptides.


Help!!!

Help!!! - ELISA Assay Forum

Help!!! - ELISA Forum. Discuss and post questions including optimization, troubleshooting, protocols and theory of ELISA assays using monoclonal, and polyclonal antibodies or peptides.


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  #1  
Old 07-20-2012, 04:43 PM
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Default Help!!!



I am with an Elisa for detect activin A, we should let it overnight with the capture antibody at room temperature but i let it at 4C, should I wait 12 hours letting it at room temperature or what shoud i do? somebody told me that I could put it at 37C 1 hour, that wont affect the assay?
Please help!!!
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Old 07-21-2012, 03:37 AM
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Default Re: Help!!!

It will probably benefit from some time at RT, at worst you ODs might be a little lower than otherwise. Which kit are you using?
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Old 07-22-2012, 06:44 PM
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Default Re: Help!!!

Isn’t important that you possess a lot ideas, but that you live one of it. Each researcher must titrate a particular antigen to obtain standardized coating conditions. If your antibody anti-activin A is polyclonal or monoclonal (rh-FS228, 2F8:6H5, etc…) or also from chicken derived (IgY), is necessary that different operative conditions must be tried, for to obtain a detection limit, with which 95 % of the normal and pathological values of the samples are correctly identified. For Mab afore mentioned I wish to acquaint the coating optimized conditions :
Mab concentration : 4-10 g/ml
Coating buffer : sodium bicarbonate 0.2 M at pH 9.4
Incubation time : 16 h (overnight)
Incubation temperature : 25 C (room)
Shaking : optional, gently at 500-600 rpm

Good luck and to be on hand
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