Plasmid Concentration - 25ug per t75 flask
Cell number - 10 million cells
Protein size - 25 KDa
Purification procedure - His tag purification using cobalt resin.
I am going to use this purified protein for crystallography studies and functional studies as well. I need to know rough yield of purified protein to calculate the number of transfection reaction
Dear Balakumaran Sathiyamani
is it very difficult predict the protein yield since it depends a lot from protein folding, stability, signal peptide.
I expressed several soluble domain of human antigeb in Expi293 (this is an improved version of HEK293 able to growth up to high cell density) an in some lucky cases i obtained very high yiedls (eh 10mg fro 25ml) while in other cases very poor yields (0,1-0,2mg from the same volume).
in the following link you can find some more information and examples about result obtained with Expi293
https://www.blogger.com/video.g?token=AD6v5dyTuEwDXD2ztCnLIFHkG06cCp1jaRuUHOGFtHdiTFnj-yS_TyCyPqzvi6RogJaSP_sXqbJnMDIYplkXODpfOc21BETPPh74J7FJuwMPxHW4X79mURcAl7-V0ApA8BQrGUAgcSc
best
Manuele
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