Cell surface differentiation marker analysis using flow cytometry

FI Fatin Najiah Mohd Idrus
NA Nurul Shuhadah Ahmad
CH Chee Hock Hoe
MA Maryam Azlan
FN Farisha Alia Norfuad
ZY Zurkurnai Yusof
WI Wan Yus Haniff Wan Isa
AA Akbar Ali Mohamed Ali
GY Get Bee Yvonne-Tee
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Polarized M1 and M2 macrophages were detached from tissue culture flask using Accumax™ (Nacalai Tesque, Japan). 2 × 105/ml M1 and M2 macrophages were washed with PBS and stained using antibodies for CD11b-PE, CD14-FITC, CD86-FITC, *CD80-APC/H7, CD206-APC and CD200R-PE (Miltenyi Biotec, Germany) (*BD Biosciences, United States) for 10 min at 4 °C in the dark. Antibodies with similar conjugates were stained in separate tubes. The cells were washed with MACS buffer to remove excess antibodies. Fluorescence was measured via flow cytometry using a FACSCanto™ II flow cytometer and FACSDiva software (BD Biosciences, United States). Histograms were plotted using FlowJo™ version 10 software (BD Biosciences, United States).

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