University:
The Pennsylvania State University
Course:
BMB 443W | Laboratory in Protein Purification and Enzymology
Academic year:

2024
Views:

200

Pages:

2
Author:

Alexis H.

Cibacron Cibacron blue blue The specific interaction between the desired molecule and the ligand attached to the chromatographic column matrix helps in purifying biological compounds Blue sepharose 6 fast flow is a type of chromatography medium consisting of cibacron blue 3G linked to the matrix by the triazine coupling method, giving a highly stable resin with minimal nonspecific adsorption chromophore matrix Structure of cibacron blue and its linkage to the matrix soH HN so Me so Elution of LDH from the affinity column Since the bonds stabilizing LDH binding to Cibacron Blue are noncovalent and electrostatic in nature, increasing the ionic strength (i.e., NaCl concentration) should weaken the binding affinity and cause LDH to elute from the column The drawback to this method is that other proteins that have bound to Cibacron Blue are also likely to dissociate under similar conditions, and so the resulting LDH is not completely purified A more selective method to elute LDH is to add the natural ligand for the active site, in this case NADH Aside from expense, the only drawback to NADH as the eluting compound is that other NADH-requiring enzymes that have bound to the column will elute along with LDH

Cibacron Blue