Technique / Immunology / ELISA assay


ELISA assay introduction



ELISA assay introduction

Introduction to ELISA assay including:

1. Sandwich ELISA Protocol
2. Competitive ELISA Protocol
3. ELISA Troubleshooting

This excellent ELISA procedure introduction is provided by Chemicon.

Enzyme-linked Immunosorbent Assays (ELISAs) combine the specificity of antibodies with the sensitivity of simple enzyme assays, by using antibodies or antigens coupled to an easily assayed enzyme that possesses a high turnover number. ELISAs can provide a useful measurement of antigen or antibody concentration.

Last update 04-Mar-2001, Rating of 7 votes.

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By Raha Favaedi on 09-Dec-2008
I'm going to set up an indirect ELISA, I have to prepare a serial dilution of antigen.
Is there both negative control & blank in indirect ELISA?

What is the difference between blank and negative control ? How can I determine threshold of positive results(450nm)?Is there any reference that describes dilutions?

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By saurabh on 27-Sep-2008
i want to known that one antibody bind with another antibody in indirect ELISA, why?
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By paro on 19-Aug-2008
it's really helpfull
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By dereje on 03-Jun-2008
is there any urine antibody test? what is the advantege?
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By muznah khatoo on 31-Oct-2006
This forum is great. All my doubts have been answered except for one. In sandwich ELISA, why do enzymes such as HRPO and chromogen/substrate solution added from right to left of the titre plate.? I hope to be given an answer and full explanation regarding to the answer of this question. Thank you

Overall good forum
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By preeti on 20-Jun-2006
My question is related to ELISA for polysaccharide vaccine.Can u suggest some of the blocking agents or treatments to evade non specificity in ELISA for polysaccharide vaccines?
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By preeti on 20-Jun-2006
My question is related to ELISA for polysaccharide vaccine.Can u suggest some of the blocking agents or treatments to evade non specificity in ELISA for polysaccharide vaccines?
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By R.P.Tiwari on 22-May-2004
We are developing a ELISA based HCV Kit, in which various blocking agents have been tried,but we are getting high background, ranging from 0.15-3.2.please suggest us ,how this can be resolved.

Till we have used, BSA, Chick albumin, PVP, Casein digest,Milk protein, Horse serum, Rabbit serum,etc
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By sa on 22-May-2004
sd
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By lynn xiao on 20-Mar-2004
Are there any other blocking agents except BSA, casein,milk?
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By MMC on 04-Mar-2001
My question is rather general in some respects. Would extremely stringent washing of the ELISA plates during the ELISA procedure result in breaking the bonds between the antibody-enzyme-substrate interaction?



As well, how would bubbles, left from washing with washing buffer, affect the addition and binding of the antibody/enzyme/substrate?
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