Tumor and lymph node cells isolated from mice were stained with LIVE/DEAD Fixable Violet Dead Staining Kit (Invitrogen) and labeled with a cocktail of monoclonal antibodies conjugated with fluorochromes: Anti-CD45 (30-F11) BV 605, anti-CD3 (17A2) BV 650, anti-CD4 (RM4-5) FITC, anti-CD8 (53–6.7) APC/Fire750, anti-CD25 (PC61) PE, anti-CD44 (IM7) PE-Cy7, anti-CD62L (MEL-14) PerCP-Cy5.5 (all from BioLegend), and anti-CD49b (DX5) PE-CF594 (BD Biosciences) for lymphoid cell analysis in tumors and anti-CD4 (RM4-5) PerCP-Cy5.5, anti-CD8 (53-6.7) PE-Cy, anti-CD44 (IM7) FITC, and anti-CD62L (MEL-14) BV 605 (all from BioLegend) for lymphocyte analysis in tLNs. In the following step, the cells were fixed using eBioscience Foxp3/Transcription Factor Staining Buffer Set (Invitrogen). Subsequently, cells from tLNs were labeled with anti-Ki67 (16A8) APC (BioLegend). Analyses of lymphoid cells in tumors and tLNs were performed using LSRFortessa (BD Biosciences).
Anti cd4 rm4 5 percp cy5
Manufactured by BioLegend
Sourced in United States
Anti-CD4 (RM4-5) PerCP-Cy5.5 is a fluorochrome-conjugated monoclonal antibody that binds to the CD4 protein on the surface of T helper cells. It can be used for the identification and enumeration of CD4+ T cells in flow cytometric analysis.
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Lab products found in correlation
2 protocols using anti cd4 rm4 5 percp cy5
1
Multiparametric Flow Cytometry of Tumor-Infiltrating Lymphocytes
2
Detailed Murine Lung and Spleen Immune Profiling
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Single-cell suspensions of the lungs and spleens were prepared as previously described30 (link). Briefly, single-cell suspensions (1 × 106 cells) from infected or immunized mice were stimulated with ESAT6 (2 μg/ml) or HEHR (2 μg/ml) at 37 °C for 9 h in the presence of GolgiPlug and GolgiStop (BD Biosciences). For phenotypical analysis, single-cell suspensions were stained with the following antibodies; Thermo Fisher Scientific (Waltham, MA, USA): Live/Dead Fixable Viability Dye eFluorTM 780; Biolegend: anti-CD90.2 (53-2.1)-BV605, anti-CD44 (IM7)-PE-Cy7, anti-CXCR3 (CXCR3-173)-APC, anti-KLRG1 (2F1/KLRG1)-BV421; BD Biosciences: anti-CD4 (RM4-5)-PerCP-Cy5.5; Biolegend: anti-CD8a (53-6.7)-BV785, anti-CD62L (MEL-14)-Alexa700, anti-CD44 (IM7)-BV421. Then, cells were fixed, permeabilized, and stained intracellularly with following antibodies; BD Biosciences: anti-IFN-γ (XMG1.2)-PE, anti-IL-2 (JES6-5H4)-PE-Cy7, anti- TNF-α (MP6-XT22)-APC, anti-IL-17A (TC11-18H10)-Alexa488, anti-IL-17A (TC11-18H10)-APC-Cy7.
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