TSKgel Chelate-5PW
TSKgel Chelate-5PW is based on a 100 nm (1000 Å) pore size polymethacrylate base material covalentely bonded with iminodiacetic acid (IDA) groups. Prior to chromatography, a metal ion, such as Zn2+, Ni2+ or Cu2+, is added to form a chelate with the IDA group. Peptides and proteins containing histidine residues will normally adsorb to these chelated ions at neutral pH. The retained compounds can be eluted with buffer containing imidazole or glycine.
Structure of TSKgel Chelate-5PW |
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Analysis of yeast enzymes |
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Column: TSKgel Chelate-5PW, 10 µm, 8 mm ID × 7.5 cm, glass
Metal ions: Zn2+, saturated
Mobile phase: 40 min linear pH gradient from 20 mmol/L
HEPES-MES-acetic acid, pH 8.0, to 20 mmol/L
HEPES-MES-acetic acid, pH 4.0, both in 0.5 mol/L NaCl
Flow rate: 1.0 mL/min
Detection: UV @ 280 nm
Recovery: G6PDH*: 90%, and ADH*: 97% of enzymatic activity
Sample: yeast enzyme concentrate
Purification: G6PDH: 8.7-fold, and ADH: 3.9-fold
*G6PDH: glucose-6-phosphate dehydrogenase
*ADH: alcohol dehydrogenase |