Quantitative analysis of pulmonary surfactant proteins B and C.
A procedure for the quantification of surfactant protein B (SP-B) and surfactant protein C (SP-C) in surfactant preparations is described. After butanol extraction of surfactant, the resulting extract is separated by Sephadex LH-60 HPLC, using an elution fluid which consists of 30% dichloromethane, 65% methanol, and 5% 100 mM HCL. SP-B and SP-C are detected by absorbance at 228 nm and quantified using standard stock solutions. Run length is approximately 70 min. Detection limits are 1 microgram SP-B and 4 micrograms SP-C.
van Eijk
M
M
Laboratory of Veterinary Biochemistry, Utrecht University, The Netherlands.
De Haas
C G
CG
Haagsman
H P
HP
D020001:1-Butanol; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D000440:Butanols; D002850:Chromatography, Gel / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003911:Dextrans; D000432:Methanol; D008752:Methylene Chloride; D011510:Proteolipids / Q000032:analysis*; D011663:Pulmonary Surfactants / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016622:Silver Staining; D012997:Solvents; D013552:Swine
D000596:Amino Acids; D000440:Butanols; D003911:Dextrans; D011510:Proteolipids; D011663:Pulmonary Surfactants; D012997:Solvents; D008752:Methylene Chloride; D020001:1-Butanol; C025614:sephadex; C040462:sephadex, 2-hydroxypropyl ether; D000432:Methanol