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Immunoblotting assay Measure the protein of the about 50 g of every sample through immunoblotting assay.Having under the situation of contrast, to carry out all processing in triplicate.Protein is separated on the 12%TRIS-HCl gel, to nitrocellulose filter, use 5% milk and TBST solution to seal, educate with a temperature resistance then through electrophoretic transfer.Resist one and in 5%BSA and TBST solution, to be incubated overnight under 4 .Resist 4 of following incubations 1 hour two.All antibody are available from CellSignaling Technology.Except that the actin uses with 1: 5000 ratio, use antibody with 1: 1000 ratio.Trace is developed, utilize densitometric analysis software I mage J quantitative result based on NIH Java.Also survey all traces, and carry out standardization to the expression of their actin separately

Nature 402 , 615622 (1999)

Wang YJ, Guan Z, Xie LY, Zhao RX, Li YY, Guan ZP & Zhang FG

What is the difference between Ipamorelin and GHRP-2 in this blend