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Anti icos

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Anti-ICOS is a laboratory reagent used in scientific research. It is an antibody that binds to the ICOS (Inducible T-cell COStimulator) protein, which is expressed on the surface of activated T cells. The function of Anti-ICOS is to detect and measure the presence of ICOS in biological samples.

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Lab products found in correlation

Anti cd4, by BioLegend (6 mentions) Anti cd19, by BioLegend (5 mentions) Streptavidin bv421, by BioLegend (4 mentions) Anti cxcr5 biotin, by BD (4 mentions) Anti foxp3, by Thermo Fisher Scientific (4 mentions) Foxp3 fix perm buffer, by Thermo Fisher Scientific (4 mentions) Anti pd 1, by BD (3 mentions) Anti cd38, by BioLegend (3 mentions) Anti cd11b, by BioLegend (3 mentions) Anti ia, by BioLegend (3 mentions) Anti ki67, by BD (3 mentions) Anti glut1, by Abcam (3 mentions) Anti cd3, by BioLegend (2 mentions) Anti gitr, by BioLegend (2 mentions) Anti hb egf dtr, by R&D Systems (2 mentions) Anti siglecf, by BD (2 mentions) Anti ige, by BD (2 mentions) Anti gl7, by BD (2 mentions) Anti cd11c, by BioLegend (2 mentions) Anti cd8a, by BioLegend (2 mentions)

Spelling variants of this product

Anti-ICOS PE (4 citations) Anti-ICOS (C398.4A, (4 citations) PE-conjugated anti-ICOS (3 citations) Anti-ICOS-PECF594 (2 citations) FITC-anti-ICOS (2 citations) -anti-ICOS-L (2 citations) Anti-ICOS (clone C398.4A; (2 citations)

9 protocols using anti icos

1

Multiparametric Immune Profiling of T-cell Subsets

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MyLa or HuT78 cell pellets were resuspended in FACS buffer (PBS, 5% FBS) and stained with predetermined antibody volumes: anti-CD107a (Cat# 555800, BD Bioscience), anti-IFNγ (Cat# 506510, Biolegend), anti-PD1 (Cat# 560908, BD Biosciences), anti-PD-L1 (Cat# 563741, Biolegend), anti-ICOS (Cat# 564778, Biolegend), anti-CTLA4 (Cat# 561717, BD Biosciences), anti-TIM3 (Cat# 565566, BD Biosciences), anti-LAG3 (Cat# 369318, BD Biosciences) and the fixation/permeabilization solution kit (Cat# 554714, BD Bioscience). Cells were analyzed using a Fortessa flow cytometer (BD Bioscience, San Jose, CA). The FlowJo 10.6.2 software program (Tree Star, Inc.) was used for data analysis.
Han Z., Estephan R.J., Wu X., Su C., Yuan Y.C., Qin H., Kil S.H., Morales C., Schmolze D., Sanchez J.F., Tian L., Yu J., Kortylewski M., Rosen S.T, & Querfeld C. (2021). MiRNA regulation of T cell exhaustion in cutaneous T cell lymphoma. The Journal of investigative dermatology, 142(3 Pt A), 603-612.e7.
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2

Multiparameter Immunophenotyping Protocol

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The following antibodies were used for surface staining at 4°C for 30 min: anti-CD4 (Biolegend, RM4-5), anti-ICOS (Biolegend, 15F9), anti-CD19 (Biolegend, 6D5), anti-PD-1 (Biolegend, RMP1-30), anti-CXCR5 biotin (BD Biosciences, 2G8), T- and B-cell activation antigen (BD Biosciences, GL-7), CD38 (Biolegend, 90), CD138 (Biolegend, 281-2), and anti-CD95 (BD Biosciences, Jo2), Streptavidin-BV421 (Biolegend, 405225). For intracellular staining, samples were fixed with the Foxp3 Fix/Perm buffer set according to the manufacturers instructions (eBioscience). Samples were then intracellularly stained with anti-FoxP3 (eBiosciences, FJK-16S). No viability dye was included. Samples were analyzed on FACS Canto II (BD) or Cytek Aurora. Data was analyzed using FlowJo v10 (FlowJo LLC).
Cavazzoni C.B., Hanson B.L., Podestà M.A., Bechu E.D., Clement R.L., Zhang H., Daccache J., Reyes-Robles T., Hett E.C., Vora K.A., Fadeyi O.O., Oslund R.C., Hazuda D.J, & Sage P.T. (2022). Follicular T cells optimize the germinal center response to SARS-CoV-2 protein vaccination in mice. Cell Reports, 38(8), 110399.
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3

Immunophenotyping and T Cell Function

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ASTRLs and PBMCs were immunophenotyped for various cell surface markers by flow cytometry with fluorophore conjugated human anti-CD3, anti-CD4, anti-CD8, anti-CD25, anti-CD127, anti-CD39 and anti-CD73 anti-CTLA4, anti-GITR, anti-ICOS, anti-CD45RA, anti-CD226, anti-LAP, anti-GARP, anti-CD56, anti-CD16, anti-CD19, anti-CD11b, anti-CD38, anti-CD27, and anti-CD24 (Biolegend). The data were acquired using a Canto II cytometer (BD Biosciences) and analyzed using Flowjo. The gating strategy for phenotyping included initial gating of a live PBMC population followed by the CD3+CD4+ population. The expression of CD25, CD127, CD39, and CD73 were expressed as % of the CD3+CD4+ population.
T cell proliferation and suppression was determined by CFSE dye dilution of the responder cells. Analysis of CFSE distribution was performed on Flowjo Proliferation platform and data are represented by Replication Index (RI). RI, determines the fold-expansion of only the responding cells, and it is the average number of divisions that all cells have undergone after they had been stained by a cell proliferation dye (13 (link)). The percentage of suppression was calculated from proliferation and suppression values.
Tripathi S., Martin-Moreno P.L., Kavalam G., Schreiber B.L., Waaga-Gasser A.M, & Chandraker A. (2022). Adenosinergic Pathway and Linked Suppression: Two Critical Suppressive Mechanisms of Human Donor Antigen Specific Regulatory T Cell Lines Expanded Post Transplant. Frontiers in Immunology, 13, 849939.
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4

Multiparameter Immune Cell Phenotyping

Cited 2 times
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The following antibodies were used for surface staining at 4°C: anti-CD4 (Biolegend, 1:200, RM4-5)12 (link), anti-ICOS (Biolegend,1:200, 15F9)12 (link), anti-CD19 (Biolegend,1:200, 6D5)12 (link), anti-CXCR5 biotin (BD Biosciences,1:100, 2G8)12 (link), GL7 (BD Biosciences,1:200, GL-7)12 (link), CD69 (Biolegend,1:200, H1.2F3)12 (link), and anti-IA (Biolegend,1:200, M5/114.15.2)12 (link). For further CXCR5 detection, streptavidin-BV421 (Biolegend, 1:400, 405225) was used at 4°C. For intracellular staining, samples were fixed with the Foxp3 Fix/Perm buffer set according to the manufacturer’s instructions (eBioscience). Samples were then intracellularly stained with anti-IgG1 (BD Biosciences, 1:200, A85-1)12 (link), anti-FoxP3 (eBiosciences, 1:200, FJK-16S)12 (link), anti-Ki67 (BD Biosciences, 1:100, B56)12 (link), anti-Glut1 (Abcam, 1:200, EPR3915)31 (link), anti-Shmt2 (Abcam, 1:200, ab64417,)32 (link) or anti-Shmt1 (Novus Biologicals, 1:200, NBP2-32173,)32 (link) at 4°C. In some cases, a donkey anti-rabbit BV421 secondary was used (Biolegend, 1:400, 406410)
Sage P.T., Ron-Harel N., Juneja V.R., Sen D.R., Maleri S., Sungnak W., Kuchroo V.K., Haining W.N., Chevrier N., Haigis M, & Sharpe A.H. (2016). Suppression by TFR cells leads to durable and selective inhibition of B cell effector functions. Nature immunology, 17(12), 1436-1446.
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5

ICOS and CXCL13 Signaling in T Cells

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Previously activated or retrovirally transduced T cells were deprived of serum for 3 h. Approximately 2 × 106 T cells were suspended in serum-free RPMI-1640 medium, incubated with purified anti-ICOS (Biolegend) or CXCL13 (PeproTech) with or without PD-L1-Fc (R&D) of indicated concentration for 30 min in 37°C. Stimulated cells were quickly spun down and lysed by 2% SDS and loading buffer and then boiled at 100 °C for 15 min. Proteins were separated by SDS-PAGE and transferred to PVDF membranes. Membranes were blocked with TBS containing 5% milk and 0.1% Tween-20. Antibodies for immunoblotting included rabbit anti–pAKT (S473), anti-AKT, anti-actin, anti-pSHP-2 (Y542), anti-SHP2 (Cell Signaling Technology). Appropriate HRP-conjugated secondary reagents were from Jackson Immunoresearch Laboratories.
Shi J., Hou S., Fang Q., Liu X., Liu X, & Qi H. (2018). PD-1 Controls Follicular T Helper Cell Positioning and Function. Immunity, 49(2), 264-274.e4.
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6

Lymphocyte Isolation and Activation

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Cell isolation and flow cytometry of lymphocytes were as described. One million CD4+ T cells were activated with plate coated anti-CD3 and anti-CD28 (Bio X Cell). After 2 days activation, T cells were removed from stimulation and rested overnight. Live cells were purified using lymphocyte isolation buffer (MP Biomedicals), followed by stimulation with plate bound anti-CD3 (2 μg/ml) and anti-ICOS (5 μg/ml; C398.4A; Biolegend) for 24 hours.
Zhou X., Qi H., Li M., Li Y., Zhu X., Amin S., Alexander M., Diadhiou C., Davidson A, & Zeng H. (2022). mTORC2 contributes to systemic autoimmunity. Immunology, 168(3), 554-568.
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7

Multi-panel Flow Cytometry Analysis

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The following antibodies were used for surface staining at 4oC for 30 minutes: anti-CD4 (Biolegend, 1:200, RM4–5), anti-ICOS (Biolegend,1:200, 15F9), anti-CD19 (Biolegend,1:200, 6D5), anti-PD-1 (1:200, RMP1–30), anti-CXCR5 biotin (BD Biosciences,1:100, 2G8), anti-GL7 (BD Biosciences,1:200, GL-7), anti-HB-EGF/DTR (RandD Systems, 1:200, AF-259-NA), anti-CD38 (Biolegend, 1:200, 90), anti-CD138 (Biolegend, 1:200, 281–2), anti-IA (Biolegend,1:200, M5/114.15.2), anti-SiglecF (BD Biosciences, 1:200, E50–2440), anti-CD8a (Biolegend, 1:200, 53–6.7), anti-CD11c (Biolegend, 1:200, N418), anti-CD11b (Biolegend, 1:200, M1/70). For CXCR5 detection, streptavidin-BV421 (Biolegend, 1:400, 405225) was used at 4oC. In some cases, anti-IgE (BD Biosciences, 1:200, R35–72) was included to block IgE bound to cell surfaces. For intracellular staining, samples were fixed with the Foxp3 Fix/Perm buffer set according to the manufacturer’s instructions (eBioscience). Samples were then intracellularly stained with anti-IgG1 (BD Biosciences, 1:200, A85–1), anti-IgE (BD Biosciences, 1:200, R35–72), anti-FoxP3 (eBiosciences, 1:200, FJK-16S), anti-Ki67 (BD Biosciences, 1:100, B56), or anti-Glut1 (Abcam, 1:200, EPR3915). In some cases, a donkey anti-goat A647 secondary was used (Invitrogen, 1:400, A-21447). See the “Life Sciences Reporting Summary” for more details.
Clement R.L., Daccache J., Mohammed M.T., Diallo A., Blazar B.R., Kuchroo V.K., Lovitch S.B., Sharpe A.H, & Sage P.T. (2019). Follicular regulatory T cells control humoral and allergic immunity by restraining early B cell responses. Nature immunology, 20(10), 1360-1371.
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8

Multi-panel Flow Cytometry Analysis

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The following antibodies were used for surface staining at 4oC for 30 minutes: anti-CD4 (Biolegend, 1:200, RM4–5), anti-ICOS (Biolegend,1:200, 15F9), anti-CD19 (Biolegend,1:200, 6D5), anti-PD-1 (1:200, RMP1–30), anti-CXCR5 biotin (BD Biosciences,1:100, 2G8), anti-GL7 (BD Biosciences,1:200, GL-7), anti-HB-EGF/DTR (RandD Systems, 1:200, AF-259-NA), anti-CD38 (Biolegend, 1:200, 90), anti-CD138 (Biolegend, 1:200, 281–2), anti-IA (Biolegend,1:200, M5/114.15.2), anti-SiglecF (BD Biosciences, 1:200, E50–2440), anti-CD8a (Biolegend, 1:200, 53–6.7), anti-CD11c (Biolegend, 1:200, N418), anti-CD11b (Biolegend, 1:200, M1/70). For CXCR5 detection, streptavidin-BV421 (Biolegend, 1:400, 405225) was used at 4oC. In some cases, anti-IgE (BD Biosciences, 1:200, R35–72) was included to block IgE bound to cell surfaces. For intracellular staining, samples were fixed with the Foxp3 Fix/Perm buffer set according to the manufacturer’s instructions (eBioscience). Samples were then intracellularly stained with anti-IgG1 (BD Biosciences, 1:200, A85–1), anti-IgE (BD Biosciences, 1:200, R35–72), anti-FoxP3 (eBiosciences, 1:200, FJK-16S), anti-Ki67 (BD Biosciences, 1:100, B56), or anti-Glut1 (Abcam, 1:200, EPR3915). In some cases, a donkey anti-goat A647 secondary was used (Invitrogen, 1:400, A-21447). See the “Life Sciences Reporting Summary” for more details.
Clement R.L., Daccache J., Mohammed M.T., Diallo A., Blazar B.R., Kuchroo V.K., Lovitch S.B., Sharpe A.H, & Sage P.T. (2019). Follicular regulatory T cells control humoral and allergic immunity by restraining early B cell responses. Nature immunology, 20(10), 1360-1371.
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9

Comprehensive Murine Regulatory T Cell Analysis

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For flow cytometry, the cell surface was stained with the following antibodies specific for mouse proteins: anti-CD4, anti-CCR6, anti-CD25, anti-CTLA-4, anti-GITR, and anti-ICOS (BioLegend). Mouse Regulatory T Cell Staining Kit (eBioscience; San Diego, CA) was used to stain the transcription factors with anti-Foxp3 (Biolegend) and anti-RORγt antibodies (BD), anti-Blimp-1 (Biolegend) and anti-Helios (Biolegend), or intracellular cytokine staining with anti-IL-17A and anti-IL-10 (BioLegend). Data were acquired using the FACSVerseFlow Cytometer (Becton Dickinson; Mountain View, CA) or the SH800 Cell Sorter (Sony, Tokyo, Japan) and analyzed with FlowJo software (Tree Star; Ashland, OR).
Furuyama K., Kondo Y., Shimizu M., Yokosawa M., Segawa S., Iizuka A., Tanimura R., Tsuboi H., Matsumoto I., & Sumida T. (2021). RORγt+Foxp3+ regulatory T cells in the regulation of autoimmune arthritis.
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