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ELISA protocol



ELISA protocol

1. To coat the ELISA plate with diluted capture antibody and incubate overnight at 4c.
2. Wash the plate wells with ddH2O, wash with PBS-Triton twice.
3. Block non-specific binding using 1% BSA/PBS and incubate for 30-60minutes at Room Temperature.
4. Wash plate. Add standards and 100uL of diluted samples to appropriate wells.
5. Incubate for 1hour at RT. Wash.
6. Add 100ul appropriate dilution of the secondary antibody conjugated with Alkaline Phosphatase (AP) or Horseradish Peroxidase (HRP) and incubate for 1 hour. Wash.
7. Add 100uL of substrate to well and incubate at RT for 1 hour. (Add stopping solution)
8. Read plates on an ELISA microplate reader.

Last update 07-Jan-2005, Rating Good of 6 votes.


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By paari on 10-Mar-2008
dear friends ?
can any one tell me how to calculate p/n ratio in elisa after reading the sample
Rating: n/a

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By Aly Kodio on 27-Feb-2008
je suis à la rcherche du protocole ELISA d'anticorps anti-pLDH
Rating: Good

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By herve on 21-May-2007
i performed an Elisa test using Biotinylated alkaline phosphatase as detection enzyme and pNPP as substrate,but i obtained always a very high background signal and no linearity depending on the concentration.could one give me some tip please?
Thanks
herve
Rating: n/a

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By LISBETHH on 02-May-2007
deseo saber o conocer comoe suna examen de la prueva de elisa su spartes su
funcioen sy como s esabe si est ainfecatdo todo eos
Rating: Very Good

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By Dr. B. Deivasigamani on 17-Dec-2006
we can use sheep red blood cells?
Rating: Very Good

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By Aashish on 07-Jan-2005
i got good information about initiating the ELISA in my lab
Rating: Very Good

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