APC anti-mouse CD68 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Mouse ears were subjected to enzymatic digestion by dispase (1 mg/ml; STEMCELL Technologies), collagenase II (2 mg/ml; PAA), and DNase I (0.8 mg/ml, Roche) in PBS for 2 h at 1400 rpm shaking and 37°C. After digestion, the samples were filtered with a 40-μm cell strainer (BD), washed with PBS, and stained with the indicated Abs.	Cell samples were analyzed with a 10-laser flow cytometer (Gallios; Beckman Coulter), and data were analyzed with the Kaluza software (version 1.2; Beckman Coulter).

Protocol tips
Mouse ears were subjected to enzymatic digestion by dispase (1 mg/ml; STEMCELL Technologies), collagenase II (2 mg/ml; PAA), and DNase I (0.8 mg/ml, Roche) in PBS for 2 h at 1400 rpm shaking and 37°C. After digestion, the samples were filtered with a 40-μm cell strainer (BD), washed with PBS, and stained with the indicated Abs.
Downstream tips
Cell samples were analyzed with a 10-laser flow cytometer (Gallios; Beckman Coulter), and data were analyzed with the Kaluza software (version 1.2; Beckman Coulter).

Publication protocol

Mouse ears were subjected to enzymatic digestion by dispase (1 mg/ml; STEMCELL Technologies), collagenase II (2 mg/ml; PAA), and DNase I (0.8 mg/ml, Roche) in PBS for 2 h at 1400 rpm shaking and 37°C. After digestion, the samples were filtered with a 40-μm cell strainer (BD), washed with PBS, and stained with the indicated Abs. The following Abs were used: anti-mouse CD45 eFluor450 (eBioscience), anti-mouse CD45 PerCP-Cy5.5 (eBioscience), anti-mouse CD11b PE-Cy7 (eBioscience), anti-mouse Ly6G FITC (BD Pharmingen), anti-mouse Ly6C PerCP-Cy5.5 (BD Pharmingen), anti-mouse F4/80 allophycocyanin (AbD Serotec), anti-mouse F4/80 PE (eBioscience), anti-mouse CD36 Alexa Fluor 488 (BioLegend), anti-mouse CD169 PE (BioLegend), anti-mouse CD68 allophycocyanin (BioLegend), anti-mouse CD64 PerCP-Cy5.5 (BioLegend), anti-mouse CD115 allophycocyanin (eBioscience), anti-mouse CD207 PE (eBioscience), anti-mouse CD11c FITC (BD Biosciences), and anti-mouse MHC class 2 eFluor450 (eBioscience). Cell samples were analyzed with a 10-laser flow cytometer (Gallios; Beckman Coulter), and data were analyzed with the Kaluza software (version 1.2; Beckman Coulter).

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