Separation of cross-linked products of rat tail tendon collagen from monomeric components by gel chromatography in the presence of Tris dodecyl sulfate at 4 degrees C.
Rat tail tendon collagen can be solubilized by a neutral aqueous solution containing 2% Tris dodecyl sulfate at 4 degrees C without being unfolded to its constituent polypeptides. The solvent at this temperature is particularly suited for gel chromatographic separation of native collagen. Thus Sepharose CL-2B gel chromatography was found to separate the monomeric collagen molecule from the products of intermolecular cross-linking. The procedure can be effectively used to follow progress of intermolecular cross-linking with aging. The monomeric collagen can be subsequently applied to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate to evaluate the mode of intramolecular cross-linking.
D000818:Animals; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003094:Collagen / Q000302:isolation & purification*; D011487:Protein Conformation; D051381:Rats; D011919:Rats, Inbred Strains; D012967:Sodium Dodecyl Sulfate; D013710:Tendons / Q000032:analysis*; D014325:Tromethamine
D014325:Tromethamine; D012967:Sodium Dodecyl Sulfate; D003094:Collagen