Direct quantification of autologous serum albumin leakage after burn injury in mice.
The current technique determining the extent of capillary leakage after injury is to measure the leakage of dye-labeled foreign albumin. A recent report, however, demonstrated that albumin leakage is dependent upon the type of fluorescent dye used for labeling. We chose to develop and test a technique for determining the extent of vascular albumin leakage after burn injury without the use of dyes. Skin and blood samples were harvested at 3h and 7 days after burn injury in mice. Total skin lysates and extracts as well as sera were analyzed for albumin leakage. Coomassie staining and Western blot analyses of skin preparations followed by the densitometric measurement revealed increased levels of albumin, suggesting that the leakage of serum albumin started within 3h after burn injury. Instead of employing dye-labeled foreign albumin, the use of SDS-polyacrylamide gel electrophoresis of tissue extracts followed by Coomassie staining will allow for a simple and direct quantification of autologous albumin leakage due to burn as well as other types of injury.
Cho
Kiho
K
Department of Surgery, Shriners Hospitals for Children Northern California, University of California at Davis, 95817, USA.
Adamson
Lee K
LK
Hobson
Kristina G
KG
Greenhalgh
David G
DG
D000818:Animals; D015153:Blotting, Western; D002056:Burns / Q000097:blood*; D003720:Densitometry; D004195:Disease Models, Animal; D005260:Female; D007202:Indicators and Reagents; D051379:Mice; D008810:Mice, Inbred C57BL; D012394:Rosaniline Dyes; D012709:Serum Albumin / Q000032:analysis*; D012720:Severity of Illness Index; D013997:Time Factors
D007202:Indicators and Reagents; D012394:Rosaniline Dyes; D012709:Serum Albumin; C048139:Coomassie blue