Application of gelatin zymography for evaluating low levels of contaminating neutrophils in red blood cell samples

被引:19
作者
Achilli, Cesare [1 ]
Ciana, Annarita [1 ]
Balduini, Cesare [1 ]
Risso, Angela [2 ]
Minetti, Giampaolo [1 ]
机构
[1] Univ Pavia, Dept Biochem, I-27100 Pavia, Italy
[2] Univ Udine, Dept Biol & Plant Protect, I-33100 Udine, Italy
关键词
D O I
10.1016/j.ab.2010.10.019
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 arti-factual 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/10(6) RBCs. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码:296 / 297
页数:2
相关论文
共 8 条
  • [1] Neutrophil granulocytes uniquely express, among human blood cells, high levels of Methionine-sulfoxide-reductase enzymes
    Achilli, Cesare
    Ciana, Annarita
    Rossi, Antonio
    Balduini, Cesare
    Minetti, Giampaolo
    [J]. JOURNAL OF LEUKOCYTE BIOLOGY, 2008, 83 (01) : 181 - 189
  • [2] BEUTLER E, 1976, J LAB CLIN MED, V88, P328
  • [3] On the association of lipid rafts to the spectrin skeleton in human erythrocytes
    Ciana, Annarita
    Achilli, Cesare
    Balduini, Cesare
    Minetti, Giampaolo
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2011, 1808 (01): : 183 - 190
  • [4] FLEMING A, 1928, BR J EXP PATHOL, V7, P281
  • [5] ELECTROPHORETIC ANALYSIS OF PLASMINOGEN ACTIVATORS IN POLYACRYLAMIDE GELS CONTAINING SODIUM DODECYL-SULFATE AND COPOLYMERIZED SUBSTRATES
    HEUSSEN, C
    DOWDLE, EB
    [J]. ANALYTICAL BIOCHEMISTRY, 1980, 102 (01) : 196 - 202
  • [6] Control of matrix metalloproteinase catalytic activity
    Ra, Hyun-Jeong
    Parks, William C.
    [J]. MATRIX BIOLOGY, 2007, 26 (08) : 587 - 596
  • [7] RETRACTED: Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells (Retracted article. See vol. 113, pg. 2370, 2009)
    Reyes, M
    Lund, T
    Lenvik, T
    Aguiar, D
    Koodie, L
    Verfaillie, CM
    [J]. BLOOD, 2001, 98 (09) : 2615 - 2625
  • [8] Zymographic techniques for the analysis of matrix metalloproteinases and their inhibitors
    Snoek-van Beurden, PAM
    Von den Hoff, JW
    [J]. BIOTECHNIQUES, 2005, 38 (01) : 73 - 83