[Preparation, purification and identification of the rabbit antibody against extracellular region of mLAIR-1].
OBJECTIVE
To express and purify the fusion protein of the extracellular region of murine leukocyte-associated Immunoglobulin-like receptor-1 (mLAIR-1) molecule with Fc fragment of human IgG and to prepare the rabbit antibody against mLAIR-1 extracellular region.


METHODS
The eukaryotic expression vector pIg/3c-mLAIR-1 was constructed. Rabbits were immunized with the fusion protein purified from the culture medium of mLAIR-1-Fc vector-transfected CHO cells with protein A column. The rabbit antibody against mLAIR-1 extracellular region were purified through Sepharose-4B affinity column coupled with mLAIR-1-Fc. The specificity of the antibody was characterized by indirect fluorescence staining and FCM analysis.


RESULTS
The mLAIR-1-Fc fusion protein was expressed and purified. The rabbit antibody against the extracellular region of mLAIR-1 molecule was obtained, which could react with both natural mLAIR-1 molecule on P388D1 cells and recombinant mLAIR-1 molecule on transfected CHO cells.


CONCLUSIONS
The rabbit antibody against the extracellular region of mLAIR-1 is prepared successfully, which may provide a useful tool for studying the structure and function of mLAIR-1.
Zhang
Yuan
Y
Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Zhuang
Ran
R
Jin
Bo-quan
BQ
Ouyang
Wei-ming
WM
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D007074:Immunoglobulin G / Q000235:genetics*; D051379:Mice; D011817:Rabbits; D011971:Receptors, Immunologic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism
D000906:Antibodies; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; C108073:leukocyte-associated immunoglobulin-like receptor 1