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Cd4 clone okt4

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The CD4 (clone OKT4) is a laboratory reagent used for the identification and enumeration of human T-helper/inducer lymphocytes. It recognizes the CD4 antigen expressed on the surface of these cells. This reagent is intended for research use only and not for use in diagnostic procedures.

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Lab products found in correlation

Lsrii flow cytometer, by BD (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Cd3 clone sp34 2, by BD (1 mentions) Perforin clone pf 344, by Mabtech (1 mentions) Cd8 clone sk1, by BioLegend (1 mentions) Cd69 clone fn50, by BioLegend (1 mentions) Granzyme b clone gb11, by Thermo Fisher Scientific (1 mentions) Cd45 clone j33, by Beckman Coulter (1 mentions) Hla dr clone ln3, by Thermo Fisher Scientific (1 mentions) Live dead fixable aqua staining kit, by Thermo Fisher Scientific (1 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (1 mentions) Lsr 2 fortessa cytometer, by BD (1 mentions) Pd 1 clone j105, by Thermo Fisher Scientific (1 mentions) Cd3 clone okt3, by Thermo Fisher Scientific (1 mentions) Cd8 clone sk1, by BD (1 mentions) Tim 3, by BioLegend (1 mentions) Pe conjugated streptavidin, by Thermo Fisher Scientific (1 mentions) Lsrfortessa, by BD (1 mentions) Accucheck counting beads, by Thermo Fisher Scientific (1 mentions) Cd8 clone rpa t8, by BD (1 mentions)

Close spelling variants of this product

CD4 (OKT4, (4 citations) Anti-CD4 antibody (clone OKT4; (2 citations) Anti-CD4 PE-Cy5.5 (clone OKT4, (2 citations)

5 protocols using cd4 clone okt4

1

PBMC Immune Phenotyping Protocol

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PBMC were thawed and divided in two 96-well plates for immune phenotyping. Surface staining was performed with the following antibodies: CD3 (clone SP34-2), from BD Biosciences, CD4 (clone OKT-4) and β7 integrin (clone FIB504) from Invitrogen, CD8 (clone SK1) and PD-1 (clone EH12.2H7) from Biolegend and LAG3 (clone P18627), and TIM3 (Clone 344823) from R&D. Intracellular staining was performed using the Foxp3 / Transcription Factor Staining Buffer Set (Ref. LTI 00-5523-00, Invitrogen) according to manufacturer’s instructions and using the following antibodies: granzyme A (clone CB9), T-bet (clone eBio4B10) and Eomes (clone WD1928) from Invitrogen and Granzyme B (clone GB-11) from Sanquin and perforin (clone Pf-344) from MabTech). Cells from the stimulation assay were characterised through surface staining using the following antibodies: CD3 (clone SP34-2), from BD Biosciences, CD4 (clone OKT-4) from Invitrogen, CD8 (clone SK1) and CD69 (clone FN50) from Biolegend (Supplementary Table 3). Samples were acquired on LSRII flow cytometer (BD Biosciences) using the DIVA Software and data was analysed using FlowJo software. Gating strategies are shown in Supplementary Figures 6–8.
Santa Cruz A., Mendes-Frias A., Azarias-da-Silva M., André S., Oliveira A.I., Pires O., Mendes M., Oliveira B., Braga M., Lopes J.R., Domingues R., Costa R., Silva L.N., Matos A.R., Ângela C., Costa P., Carvalho A., Capela C., Pedrosa J., Castro A.G., Estaquier J, & Silvestre R. (2023). Post-acute sequelae of COVID-19 is characterized by diminished peripheral CD8+β7 integrin+ T cells and anti-SARS-CoV-2 IgA response. Nature Communications, 14, 1772.
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2

Multiparameter Flow Cytometry Immunophenotyping

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The following fluorochrome-labeled mouse monoclonal antibodies were from Becton Dickinson (BD Biosciences, San Jose, CA, USA): anti-CD8 (clone RPA-T8), IFN-γ (clone: 4S.B3), IL-2 (clone MQ1-17H12), MIP-1β (clone D21-1351) and granzyme B (clone GB11); and from eBioscience (San Diego, CA): anti- CD3 (clone UCHT1), CD16 (clone CB16), CD56 (clone CMSSB), HLA-DR (clone LN3), CD38 (clone HIT2), CD69 (clone FN50), TNF-α (clone MAb11) and CD4 (clone OKT-4). In addition, we used CD45 (clone J.33) from Beckman Coulter (Fullerton, CA, USA) and perforin (clone B-D48) from BioProducts (Middletown, MD, USA).
Taborda N.A., González S.M., Alvarez C.M., Correa L.A., Montoya C.J, & Rugeles M.T. (2015). Higher Frequency of NK and CD4+ T-Cells in Mucosa and Potent Cytotoxic Response in HIV Controllers. PLoS ONE, 10(8), e0136292.
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3

Quantifying Tumor-Infiltrating T Cells

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Cells were resuspended in FACS staining buffer (PBS + 3% fetal bovine serum) using the following antibodies: CD3 (clone OKT3, eBiosciences), CD4 (clone OKT4, eBiosciences), CD8 (clone SK1, BD), PD-1 (clone J105, eBiosciences), TIM3 (clones F38–2E2, BioLegend), Tbet (clone 4B10, eBiosciences). CD19 CAR was detected using an anti-idiotype antibody provided by Novartis Pharmaceuticals. All changes in overall tumor or T cell counts reflect changes in absolute cell counts, which were determined using CountBright absolute counting beads (ThermoFisher). Cell viability was established using Live/Dead Aqua fixable staining kit (ThermoFisher), and data were acquired on an LSRII Fortessa Cytometer (BD). All data analysis was performed using FlowJo 9.0 software (FlowJo, LLC).
Singh N., Lee Y.G., Shestova O., Ravikumar P., Hayer K.E., Hong S.J., Lu X.M., Pajarillo R., Agarwal S., Kuramitsu S., Orlando E.J., Mueller K.T., Good C.R., Berger S.L., Shalem O., Weitzman M.D., Frey N.V., Maude S.L., Grupp S.A., June C.H., Gill S, & Ruella M. (2020). Impaired death receptor signaling in leukemia causes antigen-independent resistance by inducing CAR T cell dysfunction. Cancer discovery, 10(4), 552-567.
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4

HCMV-specific T cell detection

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Cells were counted by flow cytometric analysis using Accucheck counting beads (Life Technologies) and propidium iodide for exclusion of dead cells. Analysis of the T cell phenotype was performed using the following antibodies: CD3 (clone SK7, BD biosciences), CD4 (clone OKT4, eBioscience), CD8 (clone RPA-T8, BD biosciences), CD56 (clone NCAM1.2, BD biosciences). The gB-BiTE antibody construct was detected via the His-tag using an anti-His antibody (clone GG11-8F3.5.1, Miltenyi Biotec). Expression of the gB-CAR in T cells was detected with a biotinylated anti-human IgG mab (clone JDC-10, Southern Biotec) and subsequent staining with PE-conjugated streptavidin (eBioscience). Staining of HCMV-gB was performed with an antibody obtained from supernatants of the hybridoma cell line gB-27-28717 (link), which also served for construction of the scFv used in the CAR and the BiTE antibody construct. Binding of this antibody was detected by secondary staining with a PE-conjugated anti-mouse antibody (eBioscience). Non-infected and HCMV-infected HFF were blocked before antibody staining by pre-incubation with 10% human serum from an HCMV-seronegative individual (10 min at 4 °C) in order to avoid unspecific antibody binding to HCMV-encoded Fc receptors. Flow cytometric analysis was performed using a BD LSR Fortessa (BD Biosciences) and FlowJo software (FlowJo Llc.) for data analysis.
Brey C.U., Proff J., Teufert N., Salzer B., Brozy J., Münz M., Pendzialek J., Ensser A., Holter W, & Lehner M. (2018). A gB/CD3 bispecific BiTE antibody construct for targeting Human Cytomegalovirus-infected cells. Scientific Reports, 8, 17453.
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5

Detailed Immunophenotyping of B-cell Subsets

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Anti-human CD38 (clone HIT2), CD19 (clone SJ25C1), HLA-DR (clone G46-6), CD45RA (clone HI100), CXCR5 biotin (clone RF8B2), PD-1 (clone EH12.1), IL-7R (clone HIL-7R-M21), Bcl6 (clone K112-91), IgM (clone H1-FB1), CXCR3 (clone IC6), CD57 (clone NK-1), PD-1 (clone EH12.1) CXCR3 (clone 1C6/CXCR3), and CD27 (clone M-T271), all BD Biosciences. Anti-human IgD (clone IA6-2), CCR7 (clone 3D12), CCR6 (clone R6H1), CD27 (clone O323) and CD4 (clone OKT4), all eBioscience. Anti-human CD24 (clone ML5), ICOS (clone C398.4A), Foxp3 (clone 259D), CD25 (clone M-A251), streptavidin brilliant violet 605, CD4 (clone OKT4), CXCR5 (clone J252D4), CD3 (clone OKT3), CD19 (clone HIB19), and DAPI, all BioLegend. Invitrogen near-IR fixable live-dead stain kit.
Wallin E.F., Hill D.L., Linterman M.A, & Wood K.J. (2018). The Calcineurin Inhibitor Tacrolimus Specifically Suppresses Human T Follicular Helper Cells. Frontiers in Immunology, 9, 1184.
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