title: Calibrating gelatin zymograms with human gelatinase standards.
abstract: We describe the use of gelatinase standards from human capillary whole blood for calibrating gelatin zymograms. Capillary blood was obtained by fingerstick puncture and prepared in nonreducing Laemmli SDS-PAGE sample buffer without heating. Gelatin zymography revealed that whole blood (0.25- 1 microliter) contained substantial fibroblast-derived (72-kDa) and neutrophil-derived (92-, 130-, and 225-kDa) gelatinases which could be used for calibration purposes. Calibration was linear under different electrophoretic conditions using 6.5-10% polyacrylamide gels containing 1-2 mg/ml gelatin (correlation range, r = 0.990-0.998; perfect correlation, r = 1.000). The use of the standards for characterizing gelatinases from arthritic joint fluid is demonstrated. The standards are easily obtained and well characterized, and should facilitate interlaboratory comparison and standardization.
authors.affiliation: Department of Laboratory Medicine, University of Connecticut, School of Medicine, Farmington 06030, USA.
authors.forename: G S
authors.initials: GS
authors.lastname: Makowski
authors.forename: M L
authors.initials: ML
authors.lastname: Ramsby
mesh_terms: D001168:Arthritis / Q000201:enzymology; D002138:Calibration; D002196:Capillaries; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000201:enzymology; D005780:Gelatin; D018093:Gelatinases; D006801:Humans; D007962:Leukocytes / Q000201:enzymology; D008666:Metalloendopeptidases / Q000097:blood*; D012015:Reference Standards; D013582:Synovial Fluid / Q000201:enzymology
chemical_list: D005780:Gelatin; D018093:Gelatinases; D008666:Metalloendopeptidases