Aim: 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.
Conclusion: 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.