CD200 antibody | OX-90

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Cells were incubated with antibodies in 0.1 M PBS (pH7.4) supplemented with 1% FCS and 0.1% sodium azide on ice for 30 minutes. Cells were then washed three times and fixed in 1% paraformaldehyde followed by flow cytometry analysis.	Data were analyzed using the flowjo software (Tree Star, Inc., OR).

Protocol tips
Cells were incubated with antibodies in 0.1 M PBS (pH7.4) supplemented with 1% FCS and 0.1% sodium azide on ice for 30 minutes. Cells were then washed three times and fixed in 1% paraformaldehyde followed by flow cytometry analysis.
Downstream tips
Data were analyzed using the flowjo software (Tree Star, Inc., OR).

Publication protocol

For CD200 and CD200R staining, PE-labeled anti-CD200 (clone OX-90) and FITC-labeled anti-CD200R (OX-110) antibodies (Serotech) were used. FITC-, PE-, APC- or PercP- labeled antibodies to CD4, CD8α, CD11b, Gr1, Ly6G, Ly6C, DX5, IFN-γ, TNF-α, FoxP3 and isotype-matched control antibodies were purchased from BD Biosciences. Cells were incubated with antibodies in 0.1 M PBS (pH7.4) supplemented with 1% FCS and 0.1% sodium azide on ice for 30 minutes. Cells were then washed three times and fixed in 1% paraformaldehyde followed by flow cytometry analysis. Intracellular cytokine staining procedure was the same as we described (7). Data were analyzed using the flowjo software (Tree Star, Inc., OR).

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