Cd4 fitc clone rm4 4

Manufactured by Thermo Fisher Scientific

Sourced in Germany

CD4-FITC clone RM4-4 is a fluorescently labeled antibody used for the detection and analysis of CD4+ T cells in flow cytometry applications. The FITC fluorescent label allows for the visualization and quantification of CD4+ cells within a sample.

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Lab products found in correlation

Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (1 mentions) F4 80 apc, by BD (1 mentions) Siglecf pe clone es22 10d8, by Miltenyi Biotec (1 mentions) Anti mouse cd16 cd32 clone 93, by Thermo Fisher Scientific (1 mentions) Cd11b, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Cd3 bv421 clone 17a2, by BioLegend (1 mentions) Accucheck counting beads, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Software, by FlowJo (1 mentions)

2 protocols using cd4 fitc clone rm4 4

Immune cell identification protocol

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The following antibodies were used for identification of immune cells: anti-mouse CD16/CD32 clone 93 (eBioscience), Gr1-FITC (Ly-66) clone RB6-8C5 (eBioscience), SiglecF-PE clone ES22-10D8 (Miltenyi Biotec, Bergisch Gladbach, Germany), CD11b–peridinin-chlorophyll-protein complex (PerCP)–Cy5.5 (eBioscience), F4/80-APC (BD Biosciences), CD8a-PerCp-Cy5.5 clone 53-6.7 (eBioscience), CD4-FITC clone RM4-4 (eBioscience), MHCII clone M5/114.15.2 FITC (eBioscience), CD11c PE-Cy7 clone HL3 (eBioscience), and LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit (Thermo Fisher Scientific). Cells were stained for 30 min at room temperature and washed with PBS; LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit (Thermo Fisher Scientific) was used for 10 min to exclude dead cells. Cells were washed and analyzed by using flow cytometry with the FACS Canto II (BD Biosciences).

Teufelberger A.R., Van Nevel S., Hulpiau P., Nordengrün M., Savvides S.N., De Graeve S., Akula S., Holtappels G., De Ruyck N., Declercq W., Vandenabeele P., Hellman L., Bröker B.M., Krysko D.V., Bachert C, & Krysko O. (2020). Mouse Strain-Dependent Difference Toward the Staphylococcus aureus Allergen Serine Protease-Like Protein D Reveals a Novel Regulator of IL-33. Frontiers in Immunology, 11, 582044.

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Multiparameter Flow Cytometry of Immune Cells

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For flow cytometric analyzes, recovered immune cells from processed blood and organs were first stained for surface markers using the following anti-mouse antibodies: CD3 BV421 (clone 17A2, BioLegend), CD4 FITC (clone RM4-4, eBioscience), CD8a BV605 (clone 53–6.7, BioLegend), and/or CD44 AF700 (clone IM7, BioLegend). Live/dead cell discrimination was done using LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Thermo Fisher Scientific). Cells were then fixed followed by staining with anti-mouse FOXP3 AF700 or APC (clone FJK-16s, eBioscience). For blood samples, 25 uL of AccuCheck Counting Beads (Thermo Fisher Scientific) were added to determine absolute cell counts. Samples were acquired on an LSR II Flow Cytometer (Becton Dickinson). Flow cytometry data were analyzed using FlowJo software (FlowJo, LLC). Lymphocytes were gated on using side vs. forward scatter plots followed by doublet and live/dead discrimination. CD8+ and CD4+ T cells were then gated on live CD3+ T cells. Regulatory T cells (Treg) and effector CD8+ T cells (CD8_eff) were defined as CD4+ FOXP3+ and CD8+ CD44^hi cells, respectively. Fluorescence Minus One (FMO) controls were used to guide FOXP3 and CD44 gating strategies. Absolute cell counts were calculated using the counting beads according to the manufacturer’s protocol.

Maxwell R., Luksik A.S., Garzon-Muvdi T., Hung A.L., Kim E.S., Wu A., Xia Y., Belcaid Z., Gorelick N., Choi J., Theodros D., Jackson C.M., Mathios D., Ye X., Tran P.T., Redmond K.J., Brem H., Pardoll D.M., Kleinberg L.R, & Lim M. (2018). Contrasting impact of corticosteroids on anti-PD-1 immunotherapy efficacy for tumor histologies located within or outside the central nervous system. Oncoimmunology, 7(12), e1500108.

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