Cd4 fitc clone gk1

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CD4-FITC (clone GK1.5) is a fluorescently-labeled antibody that specifically binds to the CD4 molecule. CD4 is a cell surface glycoprotein expressed on a subset of T cells and plays a key role in the immune system.

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CD4-FITC (139 citations) CD4 (GK1.5, (44 citations) CD4 (clone GK1.5, (42 citations) Clone GK1.5 (14 citations) Anti-CD4 FITC (clone GK1.5, (7 citations) CD4-FITC (GK1.5, (3 citations) FITC-conjugated anti-CD4 (clone GK1.5; (3 citations) FITC-CD4 (clone GK1.5). (2 citations)

3 protocols using cd4 fitc clone gk1

Multicolor Flow Cytometry Analysis of Gastric Immune Cells

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FACS was performed as described previously (10 (link)), using FACSAria III (BD, Franklin Lakes, NJ). Live cells were gated using LIVE/DEAD Aqua Stain (cat #L34957; Life Technologies, Grand Island, NY). For the quantification of immune cell frequencies within the gastric mucosae, the cells were stained with the following antibodies: (i) for myeloid cells: CD11b-eFluor 450 (clone M1/70, cat #48-0112-82; eBioscience, San Diego, CA) and Ly6G-PE (clone 1A8, cat. #127607; BioLegend, San Diego, CA); (ii) for T cells: CD4-FITC (clone GK1.5, cat. #11-0041-85, eBioscience) and CD8-PerCpCy5.5 (clone 53-6.7, cat. #45-0081, eBioscience); (iii) for Natural Killer cells: NK1.1-PECy5 (clone PK136, BioLegend); and (iv) for B cells: B220-PE-Cy7 (clone RA3-6B2, cat. #103221, BioLegend) and IgM-PE (clone eB121-15F9, cat. #12-5890, eBioscience).
For the characterization of gastric myeloid immune cell overlap using 7-color FACS, dissociated gastric cells were stained with: (i) LIVE/DEAD Aqua (cat #L34957; Life Technologies), (ii) CD11b-eFluor 450 (clone M1/70, cat #48-0112-82; eBioscience), (iii) Ly6G-PE (clone 1A8, cat. #127607; BioLegend), (iv) Ly6C-APC-Cy7 (clone HK1.4, cat #128025, BioLegend), (v) F4/80-PerCpCy5.5 (clone BM8, cat #45-4801-80, eBioscience), (vi) CD103-APC (clone 2E7, cat #17-1031-82, eBioscience), and (vii) CD11c-PeCy5 (clone N418, cat #15-0114-81, eBioscience).

Kao K.D., Grasberger H, & El-Zaatari M. (2023). The Cxcr2+ subset of the S100a8+ gastric granylocytic myeloid-derived suppressor cell population (G-MDSC) regulates gastric pathology. Frontiers in Immunology, 14, 1147695.

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Isolation and Characterization of Intestinal Intraepithelial Lymphocytes

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Isolated SI IELs were washed in FACS buffer (PBS/3% FCS, VWR/PanBiotech). FACS buffer also served as staining solution upon adding 7AAD-PerCP-Cy5.5 (Biolegend) and the following fluorochrome-coupled antibodies directed against the indicated antigens for 15 min at 4 °C protected from light: EpCAM (biotinylated clone G8, Biolegend); T cell receptor beta chain (Pacific blue, clone H57-597, Biolegend); T cell receptor gamma delta chain (APC, clone GL3, Biolegend); CD4 (FITC, clone GK1.5, Biolegend); and CD8alpha (APC-Cy7, clone 53-6.7, Biolegend). After washing, cells were stained with diluted Streptavidin-PE (Biolegend). After extensive washing, cells were resuspended in FACS buffer (VWR/PanBiotech) and analyzed with the BD instrument Fortesssa.
The data were analyzed using Flowjo Software version 10.8.1. The following gating strategy was utilized: first of all, doublets were excluded and then only live, EpCAM -singlets were further analyzed. Here, TCRαβ + T cells were identified and further evaluated for CD4 + vs. CD8α + expression profiles.

Guliev R.R., Vogler T., Arend N., Eiserloh S., Wiede A., Kunert T., Dinkel M., Popp J., Schie I.W., Hildner K, & Neugebauer U. (2023). High-throughput Raman spectroscopy allows ex vivo characterization of murine small intestinal intra-epithelial lymphocytes (IEL). The Analyst, 148(9).

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Multiparametric Flow Cytometry Analysis

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The following antibodies were used: CD4 FITC (clone GK1.5, BioLegend # 100406, PRID:AB_312691), CD8α PB (clone 5H10, Thermo Fisher Scientific # MCD0828, RRID:AB_10372364), PE Hamster Anti-Mouse Bcl-2 Set (Bcl-2 clone 3F11, American hamster IgG clone A19-3, BD Biosciences # 556537, PRID:AB_396457), cleaved caspase-3 (Cell Signaling # 9661, PRID:AB_2341188), CD44 FITC (clone IM7, BioLegend # 103006, PRID:AB_312957), CD122 biotin (clone TM-β1, BioLegend # 123206, PRID:AB_940609), BrdU Alexa Fluor 647 (clone 3D4, BioLegend # 364107, RRID:AB_2566451), Bim (BD Biosciences # 559685, PRID:AB_397305), Fas PE (clone SA367H8, BioLegend # 152607, PRID:AB_2632903), TNFR1 PE (clone 55R-286, BioLegend # 113003, PRID:AB_313532), CD45.2 FITC (clone 104, BD Pharmingen # 553772, RRID:AB_1727491), anti-Stat5 (pY694) (BD Biosciences # 612599, RRID:AB_399882), F(ab′)2-goat anti-rabbit IgG secondary Ab Alexa Fluor 647 (Thermo Fisher Scientific # A-21246, RRID:AB_2535814), Streptavidin Alexa Fluor 647 (BioLegend # 405237, RRID:AB_2336066). For intracellular staining, live cells were first stained for surface proteins and then fixed, permeabilized, and stained for intracellular proteins using the Foxp3 Transcription Factor Staining Buffer Set (eBioscience) according to the manufacturer's instructions. All flow cytometry data were acquired on a BD Fortessa and analyzed with FlowJo software (TreeStar).

Pobezinskaya E.L., Wells A.C., Angelou C.C., Fagerberg E., Aral E., Iverson E., Kimura M.Y, & Pobezinsky L.A. (2019). Survival of Naïve T Cells Requires the Expression of Let-7 miRNAs. Frontiers in Immunology, 10, 955.

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