[Preparation and characterization of monoclonal antibody against human LSECtin].
OBJECTIVE
To prepare and characterize monoclonal antibody (mAb) against human LSECtin (liver and lymph node sinusoidal endothelial cell C-type lectin) protein.


METHODS
BALB/c mice were immunized with prokaryotically expressed human LSECtin protein. The splenocytes from the immunized mice were fused with murine myeloma cells (Sp2/0) and then the mAb-positive hybridoma cells were screened by indirect ELISA. Reaction of mAb to LSECtin antigen was characterized by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM.


RESULTS
Eight hybridoma cells secreting mAbs were established. The isotypes of the mAbs were IgG. Ascites titers were between 1:10(6) - 1:10(7). All the mAbs recognized human LSECtin protein on LSECtin-transfected 3T3 cells and six of the mAbs specifically recognized liver sinusoidal endothelial cells.


CONCLUSIONS
Eight anti-LSECtin mAbs have been obtained. The characterization of the mAbs indicate that they show fine specificity by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM, which can provide a powerful reagent for the functional study of LSECtin.
Du
Xue-mei
XM
Department of Immunology, Beijing Institute of Radiation Medicine, Beijing 100850, China.
Tang
Li
L
Liu
Xiao-lan
XL
Chen
Yong
Y
Gao
Jian-en
JE
He
Fu-chu
FC
Sun
Qi-hong
QH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007132:Immunoglobulin Isotypes / Q000032:analysis / Q000276:immunology; D037181:Lectins, C-Type / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; C505228:CLEC4G protein, human; D007132:Immunoglobulin Isotypes; D037181:Lectins, C-Type