Precision method to determine denaturation temperature of collagen using ultraviolet difference spectroscopy.
Ultraviolet difference spectroscopy was used to measure the thermal stability of molecular collagen under nonequilibrium conditions. A method using equipment commonly found in many laboratories and procedures for data gathering and data treatment are described. The melting temperature (Tm) was determined with high precision (S.D.: 0.10 degrees C) by means of close thermal spacing of recorded datas. Application of curve fitting procedures on the denaturation data enhanced the recognition of details in the thermal transition. Small transitions could be discriminated and thermally located with an accuracy of +/- 0.3 degrees C.
D000818:Animals; D003094:Collagen / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008297:Male; D008722:Methods; D011489:Protein Denaturation; D051381:Rats; D011919:Rats, Inbred Strains; D012867:Skin / Q000032:analysis*; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet; D013623:Tail; D013696:Temperature; D013710:Tendons / Q000032:analysis*
D003094:Collagen