Organs were freshly harvested and meshed on a 70 μm filter. Thereafter, samples were lysed with erythrocyte lysis buffer (Qiagen GmbH), washed twice with FACS buffer (3% BSA in 0.01 M PBS pH 7.2) and blocked with FcR block (Miltenyi Biotec) prior to staining. Subsequently, cells were incubated for 30 min at 4°C with the following antibodies: 3 μg/mL PE-Vio770 conjugated anti-CD3 (REA641, Miltenyi Biotec), 2 μg/mL PE anti-CD8a (53-6.7, eBioscience), 2 μg/mL PerCP anti-CD4 (RM4-5, BD Biosciences), and 3 μg/mL APC-Vio770 anti-CD19 (6D5, Miltenyi Biotec) antibodies. 1:1,000 (v/v) DAPI (SouthernBiotech) was used for discrimination of dead cells. Samples were acquired on a MACSQuant flow cytometer and analyzed with FlowJo software V10.
Percp anti cd4 rm4 5
Manufactured by BD
The PerCP anti-CD4 (RM4-5) is a fluorescently labeled antibody that binds to the CD4 surface antigen on cells. It can be used for the identification and analysis of CD4+ cells in flow cytometry applications.
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Lab products found in correlation
2 protocols using percp anti cd4 rm4 5
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Multiparametric Flow Cytometry Analysis of Immune Cells
2
Multi-parameter immune cell profiling
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Stain Kit, for 405nm excitation was purchased from Thermo Fischer Scientific. Mousespecific FITC-anti-CD45 (30-F11), eFluor450-anti-CD11b (M1/70), APC-anti-TIM3 (8B.2C12), APC-anti-CD25 (PC61.5), PerCP-eFluor710-anti-CD8 (53-6.7), PE-anti-Foxp3 (FJK-16s), and PE-Cy7-anti-CD4 (GK1.5) were purchased from eBiosciences. BV421-anti-PD-1 (29F.1A12) was purchased from BioLegend. Anti-CD16/CD32 FcBlock (2.4G2), PE-Cy7-anti-Ly6G/Ly6C (RB6-8C5), BV450-anti-CD45 (30-F11), FITC-anti-CD8 (53-6.7), PerCP-Cy5.5-anti-Ly6G/Ly6C (RB6-8C5), PerCP-anti-CD8 (53-6.7), PE-anti-CD3 (145-2C11), PE-anti-CD11b (M1/70), and PerCP-anti-CD4 (RM4-5) were purchased from BD Biosciences. Foxp3/Transcription Factor Fixation and Permeabilization Concentrate and Diluent kit (eBiosciences) was used for intracellular Foxp3 staining. Flow cytometry data were acquired using a BD Biosciences LSRII flow cytometer and BD FACSDiva software, and analyzed using FlowJo v10 software (TreeStar).
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