I need to make a lysis buffer to run ELISA on cell lysate.I have some difficulty in preparing it. I'll be appreciated if you could give me some hints esp about sodium orthovanadate.By the way, should I autoclave my buffer before adding aprotinin and leupeptin?
Here is the recipe:
1% NP-40, 20 mM Tris (pH 8.0), 137 mM NaCl , 10% glycerol, 2mM EDTA, 1mM activated sodium orthovanadate,10 mcg/mL Aprotinin,10 mcg/mL Leupeptin.
buffer for ELISA