Anti cxcr3 pe antibody

Bulk CD4⁺ T cells were isolated from spleens and CLN from young and aged B6 mice using magnetic beads according to the manufacturer’s instructions (untouched CD4⁺ T cell isolation kit, Miltenyi Biotec, Auburn, CA). The CD4 enriched cell suspensions contained ≥ 90% CD4⁺ T cells as determined by flow cytometry (data not shown).
CD4⁺CXCR3⁺ cells were isolated from the spleens and CLN from young and aged B6 mice via a two-step procedure in which CD4⁺ T cells were pre-enriched by depletion of unwanted cells using the kit described above. The CD4⁺ T cell fraction was then incubated with the anti-CXCR3_PE antibody (R&D Systems, Minneapolis, MN) and then magnetically labeled with anti-PE microbeads and positively selected for CD4⁺CXCR3⁺ cells, according to the manufacturers’ instructions (Miltenyi Biotec, Auburn, CA). These cells were used to perform adoptive transfer experiments or collected for quantitative gene expression. Young and aged bulk CD4⁺T or CD4⁺CXCR3⁺ cells (2×10⁶) were transferred intraperitoneally (i.p.) into 6–10 week old T cell deficient RAG1KO mice. Experiments were performed five weeks after adoptive transfer of bulk CD4⁺T cells or CD4⁺CXCR3⁺ cells. Eyes were collected for goblet cell density analysis and LGs were collected for histology.