APC anti-mouse CD252 (OX40L) Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		Data acquisition was performed using a flow cytometer (FACSCalibur, BD-Pharmingen, San Diego, CA, United States) by collecting a minimum of 10000 events per sample.

Downstream tips
Data acquisition was performed using a flow cytometer (FACSCalibur, BD-Pharmingen, San Diego, CA, United States) by collecting a minimum of 10000 events per sample.

Publication protocol

Data acquisition was performed using a flow cytometer (FACSCalibur, BD-Pharmingen, San Diego, CA, United States) by collecting a minimum of 10000 events per sample. The frequency of positive cells was analyzed using the program Cell Quest in two regions. The lymphocyte region was determined using granularity (SSC) and size (FSC) plot. DCs were identified as a MHC+ [CD205+, CD54 (ICAM-1)+, TLR4+, CD252 (OX40 L)+, CD256 (RANK)+ and CD254 (RANK L)+] population, and within this gate the CD11c+ population was assessed. The following mAbs were used: APC/Cy7 anti-mouse CD11c, PerCP/Cy5.5 anti-mouse CD205, FITC anti-mouse CD54, PerCP-Cy™5.5 anti-mouse I-A/I-E (MHC-II), PE-cyanine7 anti-mouse TLR4, APC anti-mouse CD252, PE anti-mouse CD256 and PE rat anti-mouse CD254 (eBioscience, San Diego, CA). Limits for the quadrant markers were always set based on negative populations and isotype controls.

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