How Do I Cut His Tags? Dialyze the protein against 20 mM Tris-HCl, pH 7.5. Determine the protein concentration. Combine 15 μg of protein and H2O (if necessary) to make a 45 μl total reaction volume. Add 5 μl of TEV Protease Reaction Buffer (10X) to make a 50 μl total reaction volume. What is
Can You Purify An Endogenous Protein Using IMAC? IMAC is a widely-used method for rapidly purifying polyhistidine affinity-tagged proteins, resulting in 100-fold enrichments in a single purification step. The chelators most commonly used as ligands for IMAC are nitrilotriacetic acid (NTA) and iminodiacetic acid (IDA). What is Ni-NTA chromatography used for? Ni-NTA Agarose is an