Armenian hamster igg apc

The following reagents were from BD Biosciences (San Jose, CA, USA) or eBiosciences (San Diego, CA, USA): Monoclonal antibodies to CD11b-FITC (M1/70), F4/80-PerCP5.5, CD11c-APC, CD3-pacific blue, CD4-FITC, CD8-APC, CXCL9-PE, CXCR3/CD183-PE and their corresponding isotype antibodies (Rat IgG2a-FITC, Rat IgG2a-PerCP5.5, Armenian hamster IgG-APC, Rat IgG2a-pacific blue, Rat IgG2a-FITC, Rat IgG2a APC, Hamster IgG-PE). Recombinant murine CXCL9 from Pepro Tech (Cranbury, NJ, USA) and purified mouse anti-CXCR3 antibodies (catalog number-155902, clone-S18001A, and lot number-B265189) was from Biolegend (San Diego, CA, USA). Liberace Cl was from Roche (Indianapolis, IN, USA). Bovine serum albumin (BSA), Gey’s balanced salt solution (GBSS), and DNase were from Sigma (St. Louis, MO, USA). Anti-CD11b, CD11c, CD4, and CD8 microbeads were from Miltenyi Biotec (Auburn, CA, USA). Diff-Quik stain set was from Dade Behring, Inc. (Newark, NJ, USA). Polycarbonate membranes, cell scraper, and Boyden chemotaxis chamber were from Neuro Probe, Inc. (Gaithersburg, MD, USA). ELISA kit for the detection of mouse CXCL9 was from R&D System (Minneapolis, MN, USA). LSRII flow cytometer from BD Biosciences (San Jose, CA, USA), FCS Express software from De Novo Software (Los Angeles, CA, USA).

BMMC were washed with PBS containing 0.5% BSA (Sigma) re-suspended in PBS containing 0.5% BSA (1.5 × 10⁶ cells/mL), and then incubated with 6 ng/mL rat anti-mouse CD117(KIT)-PE (eBioscience, San Diego, CA, United States) and 6 ng/mL Armenian hamster anti-mouse FcεRI-APC (eBioscience) for 1 h at 4°C. After washing with 0.5% BSA in PBS twice, cells were re-suspended in 100 μL 0.5% BSA/0.05% sodium azide in PBS and transferred to a round bottom 96-well plate. Cell samples were analyzed on a CytoFlex flow cytometer (Beckman Coulter, Brea, CA, United States). Rat IgG2b K -PE (eBioscience) and Armenian hamster IgG - APC (eBioscience) were used as isotype controls. Data was generated using FlowJo 10.6.2 software (Becton, Dickinson and Company, Ashland, OR, United States).