Fitc labeled anti mouse cd4 antibody

Todd Hewitt broth, yeast extract, trypticase soy broth (TSB) and Bacto^TM agar were purchased from BD Biosciences (Franklin Lakes, NJ, USA). Blood agar plates were purchased from Hanil Komed (Seongnam, Korea). 2,2,2-Tribromoethyl alcohol, 2-methyl-2-butanol, Red Blood Cell Lysis Buffer, and hematoxylin were purchased from Sigma-Aldrich Inc. (St. Louis, MO, USA). Goat anti-mouse IgG Fc-, IgG1-, IgG2b-, and IgA-HRP were purchased from Southern-Biotech (Birmingham, AL, USA). A Mouse Dendritic Cell Enrichment Set was purchased from BD Biosciences (San Diego, CA, USA). The following antibodies were purchased from BioLegend (San Diego, CA, USA): APC-labeled anti-mouse IFN-γ, APC-labeled anti-mouse IL-4, PE-labeled anti-mouse IL-17A, PE-labeled anti-mouse CD3, APC-labeled anti-mouse CD8, PerCP-labeled anti-mouse CD11c, FITC-labeled anti-mouse I-A^b for MHC class II, APC-labeled anti-mouse CXCR5, PerCP-labeled anti-mouse B220, PE-labeled anti-mouse CD69, PE-labeled anti-mouse PD-1, PE-labeled anti-mouse CD80, and FITC-labeled anti-mouse CD86 antibodies. FITC-labeled anti-mouse CD4 antibody was obtained from BD Biosciences. All of the isotypes matched with each antibody were purchased from BioLegend or BD Biosciences.

The cells were blocked with anti-CD16/32 (BD Pharmingen, Cat. No. 553141) to reduce nonspecific binding.
Antibodies for FACS included FITC-labeled anti-mouse CD4 antibody (BD Pharmingen, Cat. No. 553650), PE-labeled anti-mouse INF-γ antibody (BD Pharmingen, Cat. No. 554412), PE-Cy™7-labeled anti-mouse IL-4 antibody (BD Pharmingen, Cat. No. 560699), Alexa Fluor® 647-labeled anti-mouse IL-17A antibody (BD Pharmingen Cat. No. 560184), PE-labeled anti-mouse CD25 antibody (BD Pharmingen, Cat. No. 553075), and Alexa Fluor® 647-labeled anti-mouse Foxp3 antibody (BD Pharmingen, Cat. No. 560401).
The cells were stained with CD4, INF-γ, IL-4, and IL-17A antibodies to define Th1, Th2, and Th17 cells and CD4, CD25, and Foxp3 antibodies to define Treg cells.