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*Elution buffer contains 50mM Glutathionie in PBS(0.615g in 40mL PBS), pH to between 7 and 8. #Combine fractions with protein and dialyse O/N into 200 mL PBS in 50% glycerol (will concentrate sample ~4X)or other desired buffer#Measure protein concentration ([[Bradford Assay]] or A280[[Quantification by Absorbance at 280nm]]; concentrate if necessary and store at -20)#Analyze uninduced and induced cells (0.5 mL culture resuspended in 100 uL 2X SDS), lysate (5 uL in 2X), unbound (5 uL in 2X) and purified protein (0.5-10 ug) by SDS-PAGE
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#Pellet beads (save sample of unbound protein) and wash 3 x 50 mL cold wash buffer being careful to not suck up beads
#Pour into disposable column (BioRad # 732-6008) to collect beads. Wash with another 10 mL of PBS
#Prepare elution buffer containing 50mM Glutathionie in PBS(0.615g in 40mL PBS), adjust pH to between 7 and 8.
#Elute with ~5 mL room temp. elution buffer, collecting 10 x ~0.5 mL fractions. Check fractions for protein with Bradford assay
[[Category:Protein Purification]]
