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文献和实验Western Blotting with Alkaline Phosp
or passed several times through a 26-gauge needle.5.Centrifuge the sample for 5 minutes to pellet insoluble material, then discard pellet.6.Dilute an aliquot of the cell lysate sample at least 10-fold for the BCA (Pierce) protein concentration assay (SDS
from Biorad. Store in fridge.IV. 20% SDS (some people filter)V. 10% w/v ammonium persulfate, make fresh every month and store in fridge. Note: old bottles of dry AP can go bad.VI. N,N,N',N'-tetramethylethylene diamine (TEMED). Buy it and store in fridge.VII
Western Blotting with Alkaline Phosphatase Conjugates
viscosity, the sample may be sonicated briefly or passed several times through a 26-gauge needle. Centrifuge the sample for 5 minutes to pellet insoluble material, then discard pellet. Dilute an aliquot of the cell lysate sample at least 10
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