Generation of rat monoclonal antibodies against murine PTA1/CD226.
PTA1 (platelet and T cell antigen 1), designated as CD226, is one of the major activating receptors on NK cells. PTA1/CD226 is also broadly expressed on a variety of hematopoietic cells, including Th cells, CTLs, NK T cells, monocytes, DCs, and mast cells, and is involved in multiple immunoregulation. Murine PTA1 (mPTA1) is the homolog molecule of human PTA1. Using mPTA1-Fc as immunogen and the technique of rat B lymphocyte hybridoma, we raised five hybridoma cell lines secreting monoclonal antibodies (MAbs) to mPTA1, designated FMU-mCD226.1 approximately FMU-mCD226.5. Rat immunoglobulin class and subclass of the MAbs FMU-mCD226.1 approximately 5 were determined to be IgG2a, IgM, IgM, IgG1, and IgG1, respectively. FMU-mCD226.2, 3, 4, and 5 worked well in Western blot assay and could also recognize natural PTA1 on the surface of EL-4 cells and mPTA1 cDNA transfected 293T cells detected by FCM. Thus, successful production of rat anti-murine PTA1/CD226 monoclonal antibodies provides a new powerful tool for investigation of murine PTA1 function in the mouse model, both in vitro and in vivo.
Wang
Donglin
D
Department of Immunology, Fourth Military Medical University, Xi'an, China.
Zhang
Xinhai
X
Xu
Xiangsheng
X
Song
Chaojun
C
Xu
Zhuwei
Z
Jin
Boquan
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D005260:Female; D006801:Humans; D006825:Hybridomas; D051379:Mice; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000276:immunology; D000098862:T Lineage-Specific Activation Antigen 1
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D000098862:T Lineage-Specific Activation Antigen 1; D011993:Recombinant Fusion Proteins