Recombinant mouse gm csf carrier free

Bone marrow was harvested from 6 weeks-old C57BL/6 mice and differentiated into dendritic cells (BMDCs) using supplemented culture medium: RPMI 1640 (Life Technologies), 10% HI-FBS (Sigma-Aldrich), Recombinant Mouse GM-CSF (carrier-free) (20 ng/mL; BioLegend), 2 mM l-glutamine (Life Technologies), 1% antibiotic-antimycotic (Life Technologies), and 50 μM β-mercaptoethanol (Sigma-Aldrich). Bone marrow cells were treated with ACK lysis buffer for 10 min, washed, and plated with differentiation media at 1 million cells in 10 mL culture media. Cells were used for experiments on days 5–9.

All fluorescently tagged antibodies, αCD28/CD3 antibodies, ELISA kits, RBC lysis buffer, Foxp3/Transcription Factor Staining Buffer Set, EDTA, and Cell Proliferation Dye eFluor^™ 670 were purchased from Invitrogen. All inhibitors (modulators) were purchased from MedChemExpress. Bovine serum albumin (BSA) was purchased from VWR Life Science. HBSS, DPBS, PBS, DMEM, RPMI 1640, AIM-V medium, FBS, HI-FBS, HEPES, and non-essential amino acid solutions were purchased from Gibco. RAW-Dual cells, RAW Blue cells, QB buffer and reagent, EndoFit Ovalbumin (OVA), ODN 1826 (CpG), and all other TLR agonists were purchased from InvivoGen. Spleen Dissociation Medium and EasySep Mouse T-Cell Isolation Kit were purchased from STEMCELL Technologies. β-Mercaptoethanol was purchased from MP Biomedicals. Cell Activation Cocktail (without Brefeldin A), Recombinant Mouse GM-CSF (carrier-free) (20 ng/mL), and LEGENDplex MU Th1/Th2 Panel (8-plex) Kit were purchased from BioLegend. ProT2 MHC Class II tetramers and Pro5 MHC Class I pentamers were purchased from ProImmune. All plates, unless noted otherwise, were purchased from Thermo Fisher Scientific.