重组人CD47蛋白(ab126004)

This can sometimes happen with this product. They suggest use of 6M guanidine hydrochloride or 8M urea for it back to solution.

The protocol for incubating the antibody with the protein is the same as incubating the antibody with peptide. Ideally, you want to inucbate the antibody with a molar excess of 50-fold more protein per antibody binding site to increase the probability of complete blocking, but this becomes expensive, as you will need to use a lot of protein. You may find that less protein is sufficient, but as I said in my earlier e-mail, you risk forming conglomerates of protein and antibody which may actually increase the signal on the western blot, compared to antibody that is in not pre-incubated with protein.

I agree that the size of the band on the blot on the datasheet is lower than what is observed using other antibodies, for instance ab3283, which detects a protein at 50 kDa. I do not have an explanation, other than the possibility that Jurkat cells, the sample used to validate ab118222, have less glycosylation than CD47 of samples derived from normal tissue.