Separation of young and mature thrombocytes by a novel immuno-selection method.
Our earlier studies on the structural and functional properties of zebrafish thrombocytes have shown that they have many similarities to mammalian platelets. We have also shown that zebrafish have both young and mature thrombocytes as do mammalian platelets. In addition, we have distinguished young thrombocytes from mature thrombocytes microscopically using lipophilic DiI-C18, and have shown that young thrombocytes have higher GPIIb receptor levels. However, at present, there is no immunoselection method to separate young thrombocytes from mature thrombocytes in order to study differences among them, such as mRNA expression levels of thrombocyte specific genes. We developed a novel technique employing specific biotinylated anti-Cy3 antibody against the chromophore of DiI-C18 and using streptavidin magnetic beads to separate young thrombocytes from mature thrombocytes. Our technique separates and differentiates young and mature thrombocytes from whole blood. This method is specific and is effective with small amounts of blood.
Department of Biological Sciences, University of North Texas, 1510 Chestnut Denton, TX 76203, USA.
Vrinda
V
Kulkarni
Seongcheol
S
Kim
Lala
L
Zafreen
Pudur
P
Jagadeeswaran
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D001792:Blood Platelets / Q000737:chemistry / Q000378:metabolism*; D018189:Immunomagnetic Separation / Q000379:methods*; D008744:Methylamines / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000276:immunology; D012313:RNA / Q000302:isolation & purification; D013194:Staining and Labeling; D015027:Zebrafish / Q000378:metabolism
C422026:1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarboxyamine perchlorate; D000911:Antibodies, Monoclonal; D008744:Methylamines; D019038:Platelet Glycoprotein GPIb-IX Complex; C032193:glycoprotein receptor GPIb-IX; D012313:RNA