A simple and economical procedure for purification of muscle lactate dehydrogenase by batch dye-ligand adsorption.
A simple, economical and rapid purification procedure for muscle lactate dehydrogenase is described. It makes use of batch adsorption on Procion Red H-3B coupled to agarose through an amine linkage. The enzyme is eluted with NAD+ and sulphite, and for many purposes no more processing is necessary. Only 70 ml settled volume of adsorbent is required to process extract containing 1 g of lactate dehydrogenase, which is recovered with up to 90% yield, at over 98% purity. The method is readily scalable up or down, and makes an excellent class demonstration of affinity techniques.
O'Shannessy
K
K
School of Biochemistry, La Trobe University, Bundoora, VIC, Australia.
Scoble
J
J
Scopes
R K
RK
D000327:Adsorption; D000818:Animals; D002846:Chromatography, Affinity / Q000191:economics / Q000379:methods*; D004396:Coloring Agents / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D008024:Ligands; D009124:Muscle Proteins / Q000302:isolation & purification* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000201:enzymology*; D009243:NAD; D011485:Protein Binding; D011817:Rabbits; D012685:Sepharose; D013447:Sulfites; D013552:Swine; D014227:Triazines / Q000378:metabolism
D004396:Coloring Agents; D008024:Ligands; D009124:Muscle Proteins; D013447:Sulfites; D014227:Triazines; D009243:NAD; C011535:procion brilliant red H 3B; D012685:Sepharose; D007770:L-Lactate Dehydrogenase