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Position: Discussion forums > Bioscience biotechniques discussion forums > Immunology forum and cytometry forum
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immunocytochemistry protocol for biotinylated primary antibody
Posted by: spongya77 (IP Hidden, New member, 2)
Date: November 15, 2006 12:47PM
Hi,
This is going to be the first immunocytochemistry I perform, and I'd like to ask for advice first. I'll determine the relative levels of alpha2beta1 integrin in different human cell lines. The lab manager ordered a primary antibody that has biotin attached already. All the protocols I compiled call for a secondary antibody -evidently I'll have to change them. What difference does it make if I use a primary antibody with a biotin tag in regards to signal-to-noise ratio, sensitivity, etc? What should I use as controls, and what kind of blocking steps would be necessary? Any advice is greatly appreciated (in immunocytochemistry in general as well...). Thank you. Andras
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