Alexa fluor 647 conjugated anti mouse cd19

The peptide antigen, PLP_139-151 (PLP), was obtained from PolyPeptide Laboratories (Torrance, CA). For EAE induction incomplete Freund’s adjuvant (IFA) and killed Mycobacterium tuberculosis strain H37RA were purchased from Difco (Sparks, MD), and pertussis toxin was purchased from List Biological Laboratories (Campbell, CA). Dexamethasone (DEX) was purchased from Sigma-Aldrich (St. Louis, MO). For flow cytometry studies, Alexa Fluor® 488-conjugated anti-mouse CD3, Alexa Fluor® 647-conjugated anti-mouse CD19, and Pacific Blue™-conjugated anti-mouse CD11c, and their respective isotype controls were purchased from Biolegend (San Diego, CA). All other chemicals and reagents were analytical grade and used as received.

20 kDa 4-arm PEG-azide was purchased from JenKem Technology USA (Beijing, China). Tris(3-hydroxypropyltriazolylmethyl)amine (THPTA) and sodium ascorbate (NaAsc) were purchased from Sigma-Aldrich (St. Louis, MO). Copper (II) sulfate pentahydrate (CuSO₄ • 5 H₂O) was purchased from Acros Organics (Geel, Belgium). Alkyne functionalized peptide bearing an N-terminal 4-pentynoic acid (homopropargyl, hp) modification, hpPLP_139–151 (hp-HSLGKWLGHPDKF-OH), was obtained from Biomatik, USA, LLC (Wilmington, DE). All reagents were used as received without further purification. For in vitro cell assays and in vivo studies, female 4–6-week-old SJL/J (H-2) mice were purchased from Envigo Laboratories (Indianapolis, IN). For EAE induction, incomplete Freund’s adjuvant (IFA) and heat-killed mycobacterium tuberculosis H37RA were purchased from Difco (Sparks, MD). Additionally, pertussis toxin was purchased from List Biological Laboratories (Campbell, CA). For use in flow cytometry, TruStain fcX (anti-mouse CD16/32), R-phycoerythrin (PE)/Cy7-conjugated anti-mouse CD3, PE-conjugated anti-mouse CD86, FITC-conjugated anti-mouse CD80, AlexaFluor647-conjugated anti-mouse CD19, and BV421-conjugated anti-mouse CD11c were purchased from BioLegend (San Diego, CA).