any kind soul: need help in ELISA

[bostonman]

I have been using many commercially available ELISA kits for my research. Now I am trying to develop an ELISA assay using an antibody I have against chondroitin sulfate found in blood. Any kind soul want to give me a hand in this with regard to

How do I determinte the antibody titer? Checker board? Any elaboration on that?

How do I coat plate with the primary antibody?

Where do I buy the labelled secondary from?

How do I create the standard curve?

Any help,directions will be greatly appreciate. Thanks in advance.

[Brownie]

I guess you have found some help?

[wangerbio]

Wow, so many questions here. I think you'd better read some directions on ELISA technology first and then start your development.
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1. to determinate the antibody titer, checker-board test is always employed. and before this test, the plates should be coated with antibody or antigen (this depends on you ELISA method).
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2.to coat the antibody on the plates.
Just add the antibody or antigen into the wells and then add confining liquid. keep at 4C for 2h , rinse the plates and tap the water.
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3.to buy the secondary antibody, just find some biotech company and check their catalog;
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4.to create the standard curve :
Excel is the choice， draw scatter scatterplot-->add trend line,then you will get what you want.