Phorbal 12-myristate 13-acetate (PMA) and Ionomycin were purchased from Sigma-Aldrich. PMA was reconstituted to 10mg/ml stocks in DMSO and was further diluted to a working concentration of 40ng/μl in AIMV medium. Imiquimod, a TLR7 agonist, was purchased from LKT Laboratories (St Paul, MN) and reconstituted to 1mg/ml in DMSO. Recombinant TIMP1, TIMP2, TIMP3, and TIMP4 were purchased from R&D Systems and were reconstituted to working concentrations in AIMV medium. The protease inhibitors GM6001 and TAPI-0 were purchased from Calbiochem and reconstituted to 10mM and 1mg/ml stock, respectively, in DMSO.
The monoclonal rat anti-hCD200 antibodies 1B9 and 3G7, and the polyclonal rabbit serum against the extracellular region of CD200 (CD200v+c), were described previously [2 (link)]. A polyclonal rabbit serum against the human CD200 receptor (CD200R1) was generated by immunization of rabbits with a fusion protein containing the extracellular region of human CD200R1 with a his-tag at the N-terminal.
Antibodies against CD19 and CD62L used in FACS analyses were purchased from Biolegend. The apoptosis detection kit for staining of Annexin V and 7AAD was purchased from BD Biosciences. The Pan-Cadherin antibody, used as a plasma membrane marker for loading controls in Western blots, was purchased from Abcam.
The monoclonal rat anti-hCD200 antibodies 1B9 and 3G7, and the polyclonal rabbit serum against the extracellular region of CD200 (CD200v+c), were described previously [2 (link)]. A polyclonal rabbit serum against the human CD200 receptor (CD200R1) was generated by immunization of rabbits with a fusion protein containing the extracellular region of human CD200R1 with a his-tag at the N-terminal.
Antibodies against CD19 and CD62L used in FACS analyses were purchased from Biolegend. The apoptosis detection kit for staining of Annexin V and 7AAD was purchased from BD Biosciences. The Pan-Cadherin antibody, used as a plasma membrane marker for loading controls in Western blots, was purchased from Abcam.