Question

does cibacron blue bind to the same site as pyruvate or something else in LDH experiment?(CB was in liquid form)

Answer 1

Cibacron blue (F3G-A) or CB in this is in liquid form so it might be attached to the agarose or the water soluble Polyvinyl-alcohol(PVA) which provides inert support (so it does not bind to the same site as pyruvate in LDH experiment) and

this dye CB has the ability to mimic the pyridine nucleotides because of its charge and shape and

when the LDH experiment is performed using affinity chromatography only those protein binds with affinity column which have affinity for pyridine nucleotides and

then wash off the column first time after washing the same column second time NAD⁺ might contain with the affinity for dye and removed protein.