Hello!
I am using a protocol from Iriguchi et al. to obtain HPC and then lymphocytes.
They want us to coat plates with rhDL4/Fc protein and retronectin.
They say: "Dilute rhDL4/Fc chimera protein solution (10ug/ml) with an equal volume of Retronectin (10ug/ml) and add 150ul of the solution to each well of 48-well plates"
It is all clear but looking at the reagents I realized they have 2 different solvents - Retronectin should be dissolved in PBS while rhDL4/Fc in ddH2O.
I am not sure if it is the best way to do that. Probably retronectin coating should be made at first and then rhDL4/Fc protein should be added?
Both reagents are expensive and I don't have possibility to test every possible way of coating.
Have anyone ever used such coating combination? Or maybe do you have some tips to be sure that plates are properly coated?
Thank so much for your help!
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