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Icos clone c398.4a

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ICOS (clone C398.4A) is a monoclonal antibody that recognizes the ICOS (inducible T-cell costimulator) receptor. ICOS is expressed on activated T cells and plays a role in T cell activation and regulation.

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Lab products found in correlation

Facscanto 2 flow cytometer, by BD (2 mentions) Fixation permeabilization solution kit, by BD (2 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (2 mentions) Fixable near ir dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Igd clone 11 26, by Thermo Fisher Scientific (2 mentions) Clone okt3, by BioLegend (2 mentions) Phycoerythrin conjugated anti cd150 slamf1 clone tc15 12f12.2, by BioLegend (2 mentions) Fluorophore conjugated anti cd4, by Thermo Fisher Scientific (2 mentions) Anti mouse cxcr5, by BD (2 mentions) Phycoerythrin conjugated anti fas cd95 clone jo2, by BD (2 mentions) Allophycocyanin conjugated anti mouse bcl6 clone k112 91, by BD (2 mentions) Ikkε 2690, by Cell Signaling Technology (2 mentions) Fitc conjugated pna, by Vector Laboratories (2 mentions) Cd62l clone mel 14, by BioLegend (1 mentions) Transcription factor buffer set, by BD (1 mentions) Pd 1 clone j43, by BD (1 mentions) Triton x 100, by Merck Group (1 mentions) Fix perm buffer, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions) Sodium citrate, by BD (1 mentions)

Spelling variants of this product

ICOS (C398.4A, (8 citations) Clone C398.4A (3 citations) Anti-ICOS (clone C398.4A; (2 citations) αCD278(ICOS)-APC (clone C398.4A) (2 citations)

5 protocols using icos clone c398.4a

1

Flow Cytometry Analysis of Immune Cells

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Single-cell suspensions were washed and stained with Fixable Near-IR Dead Cell Stain Kit (Thermo Fisher, L10119) followed by staining of extracellular antigens with fluorochrome labeled antibodies. The Fixation/Permeabilization Solution Kit (BD Biosciences, 554714) was used for exposing cytoplasmic antigens. Fluorochrome-labeled antibodies against mouse antigens was used in different combinations. CD44 (clone IM7), CD45.1 (clone A20), CD45.2 (clone 104), CD8 (clone 53–6.7), CTLA-4 (clone UC10-4F10-11), were purchased from BD Biosciences. CD62L (clone MEL-14), and ICOS (clone C398.4A) were purchased from BioLegend. Anti-mouse Granzyme B (clone GB12) was purchased from Thermo Fisher. Cell counting was performed with CountBright Absolute Counting Beads (Thermo Fisher, C36950). Samples were processed in a FACSCanto II flow cytometer (BD Biosciences). Data analysis was performed with FlowJo, version 8.8.7 (Tree Star).
Rundqvist H., Veliça P., Barbieri L., Gameiro P.A., Bargiela D., Gojkovic M., Mijwel S., Reitzner S.M., Wulliman D., Ahlstedt E., Ule J., Östman A, & Johnson R.S. (2020). Cytotoxic T-cells mediate exercise-induced reductions in tumor growth. eLife, 9, e59996.
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2

Multiparametric Flow Cytometry Analysis

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Single-cell suspensions were washed and stained with Fixable Near-IR Dead Cell Stain Kit (Thermo Fisher, L10119) followed by staining of extracellular antigens with fluorochrome labelled antibodies. The Fixation/Permeabilization Solution Kit (BD Biosciences, 554714) was used for exposing cytoplasmic antigens. The Transcription Factor Buffer Set (BD Biosciences, 562574, RRID : AB_2869424) was used for exposing nuclear antigens. Fluorochrome-labelled antibodies against mouse antigens CD44 (clone IM7), CD8 (clone 53- 6.7), CTLA-4 (clone UC10-4F10-11), LAG-3 (clone C9B7W), and PD-1 (clone J43) were purchased from BD Biosciences, CD27 (clone LG.3A10), CD28 (clone 37.51), CD62L (clone MEL-14), and ICOS (clone C398.4A) were purchased from BioLegend, and 4-1BB (clone 17B5), CD127 (clone A7R34), CD25 (clone PC61.5), Eomes (clone Dan11mag), T-bet (clone eBio4B10), and anti-human/mouse Granzyme B (clone GB12) was purchased from Thermo Fisher. Cell counting was performed with CountBright Absolute Counting Beads (Thermo Fisher, C36950). Samples were processed in a FACS Canto II flow cytometer (BD Biosciences). Data analysis was performed with FlowJo (RRID : SCR_008520, version 10.7.2).
Barbieri L., Veliça P., Gameiro P.A., Cunha P.P., Foskolou I.P., Rullman E., Bargiela D., Johnson R.S, & Rundqvist H. (2023). Lactate exposure shapes the metabolic and transcriptomic profile of CD8+ T cells. Frontiers in Immunology, 14, 1101433.
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3

Monoclonal Antibodies for Immune Cell Analysis

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Monoclonal Abs specific for human CD3 (clone OKT3), and mouse CD3 (clone 145-2C11), -CD28 (clone 37.51), -CTLA-4 (clone UC10-4B9) or -ICOS (clone C398.4A) were purchased from Biolegend CA, as was phycoerythrin-conjugated anti-CD150/SLAMf1 (clone TC15-12F12.2). Fluorophore-conjugated anti-CD4 (mouse: clone RM4-5; human: clone RPA-T4), -CD8a (clone 53-6.7), -CD19 (mouse: clone eBio1D3; human: clone HIB19), -CD25 (clone PC61), -CD40L (clone MR1), -CD44 (clone IM7), -CD45.1 (clone A20), -CD62L (clone MEL-14), -GL7 (clone GL-7), -IgD (clone 11-26) and -PD1 (mouse: clone J43; human: clone J105) Abs were obtained from eBioscience, CA. Anti-mouse CXCR5 (mouse: clone 2G8; human: clone RF8B2), biotinylated anti-CD138 (clone 281-2), FITC-conjugated TCR β-chain (H57-597), phycoerythrin-conjugated anti-Fas/CD95 (clone Jo2) and allophycocyanin-conjugated anti-mouse Bcl6 (clone K112-91) were procured from BD Biosciences, CA. FITC-conjugated PNA was purchased from Vector Laboratories, CA. Monoclonal anti-TBK1 (#3013), -phospho-TBK1 (Ser172) (clone D52C2, #5483), -TRAF2 (#4712), -TRAF3 (#4729), p-ERK1/2 (T202/Y204) (clone E10, #9106) and -IKKε (#2690) Abs were obtained from Cell Signaling Technology, MA. Monoclonal anti-p85α (sc-1637), -ERK2 (sc-1647) and -TRAF5 (sc-7220) were purchased from Santa Cruz Biotechnology, CA. Recombinant IL-2 and IL-7 cytokines were obtained from Biolegend, CA.
Pedros C., Zhang Y., Hu J.K., Choi Y.S., Canonigo-Balancio A.J., Yates JR I.I.I., Altman A., Crotty S, & Kong K.F. (2016). A TRAF-like motif of ICOS controls development of germinal center T follicular helper cells via TBK1. Nature immunology, 17(7), 825-833.
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4

Multiparametric Immunophenotyping of PBMCs

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Whole blood samples were collected in cell preparation tubes with sodium citrate (BD Biosciences). PBMCs were obtained by centrifugation and viably frozen until analysis. PBMCs were thawed, washed with flow buffer (5% BSA, 2 mM EDTA in PBS) and incubated with LIVE/DEAD Fixable Aqua Dead Cell Stain (Life Technologies), Fc receptor blocking agent (Miltenyi Biotec) and stained with surface antibodies (CD3 clone OKT3, CD4 clone RPA-T4, CD8 clone SK1, CD14 clone HCD14, CD19 clone HIB19, CD25 clone BC96, ICOS clone C398.4A, HLA-DR clone L243, PD-1 clone 29F.1A12 all from BioLegend) for 20 min at 4°C. For Foxp3 and Ki67 staining, cells were fixed and permeabilized using a Fix/Perm buffer (eBiosciences) according to the manufacturer’s instructions, then stained with anti-Foxp3 (clone 206D, BioLegend) or anti-Ki67 antibody (clone B56, BD Biosciences).
For global protein acetylation analysis, after surface staining, cells were fixed with 0.4% paraformaldehyde (Thermo Fisher Scientific), permeabilized in Triton X-100 (Sigma-Aldrich) and subsequently stained with anti-acetylated lysine antibody (clone 15G10, BioLegend).
Hellmann M.D., Jänne P.A., Opyrchal M., Hafez N., Raez L.E., Gabrilovich D., Wang F., Trepel J.B., Lee M.J., Yuno A., Lee S., Brouwer S., Sankoh S., Wang L., Tamang D., Schmidt E., Meyers M.L., Ramalingam S.S., Shum E, & Ordentlich P. (2020). Entinostat plus pembrolizumab in patients with metastatic NSCLC previously treated with anti-PD-(L)1 therapy. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(4), 1019-1028.
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5

Monoclonal Antibodies for Immune Cell Analysis

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Monoclonal Abs specific for human CD3 (clone OKT3), and mouse CD3 (clone 145-2C11), -CD28 (clone 37.51), -CTLA-4 (clone UC10-4B9) or -ICOS (clone C398.4A) were purchased from Biolegend CA, as was phycoerythrin-conjugated anti-CD150/SLAMf1 (clone TC15-12F12.2). Fluorophore-conjugated anti-CD4 (mouse: clone RM4-5; human: clone RPA-T4), -CD8a (clone 53-6.7), -CD19 (mouse: clone eBio1D3; human: clone HIB19), -CD25 (clone PC61), -CD40L (clone MR1), -CD44 (clone IM7), -CD45.1 (clone A20), -CD62L (clone MEL-14), -GL7 (clone GL-7), -IgD (clone 11-26) and -PD1 (mouse: clone J43; human: clone J105) Abs were obtained from eBioscience, CA. Anti-mouse CXCR5 (mouse: clone 2G8; human: clone RF8B2), biotinylated anti-CD138 (clone 281-2), FITC-conjugated TCR β-chain (H57-597), phycoerythrin-conjugated anti-Fas/CD95 (clone Jo2) and allophycocyanin-conjugated anti-mouse Bcl6 (clone K112-91) were procured from BD Biosciences, CA. FITC-conjugated PNA was purchased from Vector Laboratories, CA. Monoclonal anti-TBK1 (#3013), -phospho-TBK1 (Ser172) (clone D52C2, #5483), -TRAF2 (#4712), -TRAF3 (#4729), p-ERK1/2 (T202/Y204) (clone E10, #9106) and -IKKε (#2690) Abs were obtained from Cell Signaling Technology, MA. Monoclonal anti-p85α (sc-1637), -ERK2 (sc-1647) and -TRAF5 (sc-7220) were purchased from Santa Cruz Biotechnology, CA. Recombinant IL-2 and IL-7 cytokines were obtained from Biolegend, CA.
Pedros C., Zhang Y., Hu J.K., Choi Y.S., Canonigo-Balancio A.J., Yates JR I.I.I., Altman A., Crotty S, & Kong K.F. (2016). A TRAF-like motif of ICOS controls development of germinal center T follicular helper cells via TBK1. Nature immunology, 17(7), 825-833.
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