Application of gelatin zymography for evaluating low levels of contaminating neutrophils in red blood cell samples.
Supposedly "homogeneous" red blood cell (RBC) samples are commonly obtained by "washing" whole blood free of plasma, platelets, and white cells with physiological solutions, a procedure that does not result, however, in sufficient removal of polymorphonuclear neutrophils (PMNs), leading to possible artifactual results. Pure RBC samples can be obtained only by leukodepletion procedures. Proposed here is a version of gelatin zymography adapted to detect matrix metalloproteinase 9 (MMP-9), selectively expressed by PMNs, in heterogeneous mixtures of RBCs and PMNs that can reveal contamination at levels as low as 1 PMN/10⁶ RBCs.
Achilli
Cesare
C
Department of Biochemistry, University of Pavia, Italy.
Ciana
Annarita
A
Balduini
Cesare
C
Risso
Angela
A
Minetti
Giampaolo
G
D002469:Cell Separation; D057075:Enzyme Assays / Q000379:methods*; D004912:Erythrocytes / Q000737:chemistry* / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism*; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology*
D005780:Gelatin; D020780:Matrix Metalloproteinase 9