Sandwish ELISA Problems and Troubleshooting

proatpcr · #1 (**permalink**) 04-13-2008, 09:20 AM

Hello, I have a big problem with my sandwich ELISAs.

I am using a mouse monoclonal antibody to coat the plate after which I block with PBS/BSA 1%.

I am using rabbit polyclonal serum against my protein of interest.
Maybe the serum polyclonal antibody contains antibodies against BSA?

The fact is that the lysis buffer has an value important and more interestingly my cells lysates without the protein of interest have the same value as 12500pg of the protein alone using the same buffer.

I do not know what i can change in the protocol. If someone can help thanks

danfive · #2 (**permalink**) 04-13-2008, 05:07 PM

Hi,
Here are some good online resources from BD biosciences:
Typical protocol: http://www.bdbiosciences.com/pharmin...gE_ELISA.shtml
Troubleshooting videos: http://www.bdbiosciences.com/pharmin...ownloads.shtml
Look at the video for Troubleshooting: high background --> http://www.bdbiosciences.com/support.../OptEIAu11.avi

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Sandwish ELISA Problems and Troubleshooting

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