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Good starting range in ELISA Capture - ELISA (Aug/01/2011 )

Hello,

I usually just use ELISA to detect antibodies in in vitro work or purified antibodies. I recently got in human serum and tried to start at 1:10. They all gave an OD > 3 which is not right. I am coating with antigen, then binding serum, then detecting for human IgG. I am currently using a 10%FBS, 5% Milk, .05% Tween as my block which works great for my purified antibodies.

I'm not sure if the block is the problem because my control antibodies worked just fine. The serum samples were just way to high.

J

-Jordan W-

Jordan W on Mon Aug 1 23:16:55 2011 said:


Hello,

I usually just use ELISA to detect antibodies in in vitro work or purified antibodies. I recently got in human serum and tried to start at 1:10. They all gave an OD > 3 which is not right. I am coating with antigen, then binding serum, then detecting for human IgG. I am currently using a 10%FBS, 5% Milk, .05% Tween as my block which works great for my purified antibodies.

I'm not sure if the block is the problem because my control antibodies worked just fine. The serum samples were just way to high.

J


Perhpas i should note that this is human plasma. Maybe I should use human serum to block? Could the plasma be binding to the FBS or milk?

-Jordan W-