Icos c398.4a

Manufactured by BioLegend

ICOS (C398.4A) is a flow cytometry antibody product from BioLegend. It is used for the detection and analysis of the inducible T-cell co-stimulator (ICOS) marker on cells.

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Lab products found in correlation

Foxp3 fjk 16s, by Thermo Fisher Scientific (3 mentions) Cd25 7d4, by BD (3 mentions) Il 17a tc11 18h10, by BD (3 mentions) Nrp1 fab566a, by R&D Systems (3 mentions) Ctla4 14d3, by Thermo Fisher Scientific (3 mentions) Gitr dta 1, by Thermo Fisher Scientific (3 mentions) Cd62l mel 14, by BioLegend (3 mentions) Cd44 im7, by BioLegend (3 mentions) Ki67 sola15, by Thermo Fisher Scientific (3 mentions) Cd4 h129, by BD (3 mentions) Pd 1 eh12.2h7, by BioLegend (2 mentions) Ab39670, by Abcam (2 mentions) Ifn g xmg1, by BD (2 mentions) Lag3 46 2231 80, by Thermo Fisher Scientific (2 mentions) P27 sc 776, by Santa Cruz Biotechnology (2 mentions) Ab85047, by Abcam (2 mentions) P foxo1 9461s, by Cell Signaling Technology (2 mentions) Ab5454, by Abcam (2 mentions) Brdu b44, by BD (2 mentions) Foxo1, by Abcam (2 mentions)

Spelling variants of this product

ICOS (clone C398.4A) (5 citations) Anti-ICOS (C398.4A, (4 citations) ICOS-PE/Cy7 (C398.4A, (2 citations) Anti-ICOS (clone C398.4A; (2 citations) αCD278(ICOS)-APC (clone C398.4A) (2 citations)

8 protocols using icos c398.4a

Innate and Adaptive Immune Phenotyping

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Monoclonal antibodies used for staining of PBMCs for innate cell phenotyping were CD3 (UCHT1, #557943), CD19 (HIB19, #557921), CD14 (M5E2, #565283), HLA-DR (G46-6, #560651), CD11c (O33-782, #561355), and CD123 (7G3,554529) from BD Biosciences (Franklin Lakes, NJ), CD56 (MEM188,17-0569) and CD16 (CB16,47-0168) from eBiosciences (San Diego, CA); for cT_FH phenotyping CD3 (SP34-2, #562877), CD4 (L200, #560836), CXCR5 (RF8B2), and CXCR3 (IC6/CXCR3) BD Biosciences, ICOS (C398.4 A) and PD-1 (EH12.2H7) from Biolegend (San Diego, CA) (Supplementary Fig. 2). Data were collected on an LSRII (BD Biosciences) and data were analyzed using R 3.6.3 software. Sample sizes available for statistical analysis differed based on sample availability (monocytes—MNP: n = 7; IM = 10, cT_FH—MNP: n = 8; IM: n = 9).

Rouphael N.G., Lai L., Tandon S., McCullough M.P., Kong Y., Kabbani S., Natrajan M.S., Xu Y., Zhu Y., Wang D., O’Shea J., Sherman A., Yu T., Henry S., McAllister D., Stadlbauer D., Khurana S., Golding H., Krammer F., Mulligan M.J, & Prausnitz M.R. (2021). Immunologic mechanisms of seasonal influenza vaccination administered by microneedle patch from a randomized phase I trial. NPJ Vaccines, 6, 89.

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Multiparameter FACS and Cell Sorting

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The following antibodies were used in fluorescence-activated cell sorting (FACS) and cell sorting: CD45.1 (A20, BioLegend), CD45.2 (104, BioLegend), CD4 (H129.19, BD Pharmingen), CD25 (7D4, BD Pharmingen), IL-17A (TC11-18H10.1, BD Pharmingen), IFN-g (XMG1.2, BD Pharmingen), CD44 (IM7, Bio-Legend), CD62L (MEL-14, BioLegend), CD25 (7D4, BD Pharmingen), Lag3 (46-2231-80, eBioscience), Nrp1 (FAB566A, R&D Systems), Ki67 (SolA15, eBioscience), CTLA4 (14D3, eBioscience), GITR (DTA-1, eBioscience), ICOS (C398.4A, BioLegend), Foxp3 (FJK-16 s, eBioscience), p27 (sc-776, Santa Cruz Biotechnology), p-p27 (ab85047, Abcam), Foxo1 (ab39670, Abcam), p-Foxo1 (9461S, Cell Signaling Technology), Raptor (ab5454, Abcam), Brdu (B44, BD Pharmingen).

Wu C., Chen Z., Xiao S., Thalhamer T., Madi A., Han T, & Kuchroo V. (2018). SGK1 Governs the Reciprocal Development of Th17 and Regulatory T Cells. Cell reports, 22(3), 653-665.

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Multiparametric Flow Cytometry Analysis of Mediastinal Lymph Nodes

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Mediastinal lymph nodes (mLNs) were harvested, and single-cell suspensions were prepared. The cells were preincubated with Fc blocker and then incubated on ice for 30 min with antibody cocktail and Fixable Viability Dye, Ghost Red 780 (Tonbo Biosciences). Flow cytometry was conducted using fluorochrome-conjugated Abs for CD3 (145-2C-11, BD Biosciences), CD4 (RM5, BD Biosciences), CXCR5 (2G8, BD Biosciences), ICOS (C398.4A, BioLegend), PD-1 (J43, BD Biosciences and 29F.1A12, BioLegend), B220 (RA3–6B2, BD Biosciences), Fas (Jo2, BD Biosciences), PNA (FL-1071, Vector Laboratories), CD11b (M1/70, BD Biosciences), Ly6G (1A8, BioLegend), and rat IgG2a isotype control (BioLegend). Data were acquired using Fortessa (BD Biosciences Immunocytometry Systems, Franklin Lakes, NJ). All data were analyzed with the FlowJo software (TreeStar, Ashland, OR).

Lama J.K., Iijima K., Kobayashi T, & Kita H. (2022). Blocking the inhibitory receptor PD-1 prevents allergic immune response and anaphylaxis in mice. The Journal of allergy and clinical immunology, 150(1), 178-191.e9.

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Immunophenotyping of EAMG Rat Immune Cells

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On day 50 (peak of the disease) after induction of EAMG, rats were euthanized under deepening anesthesia. Spleens and inguinal lymph nodes were removed and the weights were measured. Then the spleens and inguinal lymph nodes were minced through a 70 μm cell strainer to prepare single-cell suspensions of mononuclear cells (MNCs) under aseptic conditions. Red blood cells in spleen MNCs were lysed with a lysis buffer (Biolegend) for 5 min. All specimens were coded to facilitate blind testing. The spleen or inguinal lymph nodes MNCs suspension with a final volume of 100 μl were labeled with corresponding monoclonal antibodies and incubated for 30 min at 4 °C in the dark. The following antibodies were used in the assay: CD3 (1F4; Biolegend), CD4 (OX35; eBioscience), B220 (HIS24; eBioscience), CD20 (SP32; Abcam), CD161 (10/78; BD Pharmingen), MHC II (HIS19; Biolegend), CD80 (3H5; Biolegend), CD86 (24F; Biolegend), CD40 (HM40-3; Biolegend), CD27 (LG.7F9; eBioscience), ICOS (C398.4A; Biolegend), CXCR5 (ERP8837; Abcam). Unconjugated primary antibodies were detected with Alexa Fluro 488-conjugated anti-rabbit IgG (Abcam). Samples were analyzed using an Aria II flow cytometer (BD).

Zhang P., Yang C.L., Du T., Liu Y.D., Ge M.R., Li H., Liu R.T., Wang C.C., Dou Y.C, & Duan R.S. (2021). Diabetes mellitus exacerbates experimental autoimmune myasthenia gravis via modulating both adaptive and innate immunity. Journal of Neuroinflammation, 18, 244.

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Multiparameter FACS and Cell Sorting

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Wu C., Chen Z., Xiao S., Thalhamer T., Madi A., Han T, & Kuchroo V. (2018). SGK1 Governs the Reciprocal Development of Th17 and Regulatory T Cells. Cell reports, 22(3), 653-665.

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Comprehensive Immune Profiling of PBMCs

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PBMCs were stained with Fixable Viability Dye (Invitrogen) in PBS, followed by staining with fluorochrome-conjugated antibodies against CD4 (RPA-T4), CD45RA (HI100), CXCR5 (J252D4), CXCR3 (G025H7), CCR6 (11A9), PD-1 (EH12.2H7), and ICOS (C398.4A) (from BioLegend and BD Biosciences). Cells were stained in MACS buffer (0.5 % FBS, 1 % penicillin and streptomycin, and 2 mM EDTA in PBS) for 30 min at 4°C.

Kim Y.J., Oh J., Jung S., Kim C.J., Choi J., Jeon Y.K., Kim H.J., Kim J.W., Suh C.H., Lee Y., Im S.H., Crotty S, & Choi Y.S. (2023). The transcription factor Mef2d regulates B:T synapse-dependent GC-Tfh differentiation and IL-21-mediated humoral immunity. Science immunology, 8(81), eadf2248.

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Multiparametric Flow Cytometry Analysis

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Unless otherwise stated all chemicals were purchased from Sigma Aldrich. The following antibodies were used in flow cytometry (FACS) and cell sorting: CD4 (H129.19, BD Pharmingen), CD8 (53-6.7, Biolegend), CD25 (7D4, BD pharmingen), IL-4 (11B11, Biolegend), IL-17A (TC11–18H10.1, BD Pharmingen), IFN-γ (XMG1.2, BD Pharmingen), CD44 (IM7, Biolegend), CD62L (MEL-14, Biolegend), CTLA4 (14D3, eBioscience), GITR (DTA-1, eBioscience), TIGIT (GIGD7, eBioscience), ICOS (C398.4A, Biolegend), Foxp3 (FJK-16s, eBioscience), IL-10 (JES5–16E3, Biolegend), Nrp1 (FAB566A, R&D), IL-4R (I015F8, Biolegend), Ki67 (SolA15, eBioscience).

Wu C., Chen Z., Dardalhon V., Xiao S., Thalhamer T., Liao M., Madi A., Franca R.F., Han T., Oukka M, & Kuchroo V. (2017). The transcription factor musculin promotes the unidirectional development of peripheral Treg cells by suppressing the TH2 transcriptional program. Nature immunology, 18(3), 344-353.

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Multiparameter Flow Cytometry of Macaque Immune Cells

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The following fluorochrome conjugated monoclonal antibodies reactive with macaque cells were used for flow cytometry studies: CD3 (SP34-2, BD Biosciences [BD]), CD4 (L200, BD), CD8 (SK1, BD), CD95 (DX2, BD), CD28 (CD28.2, BioLegend), CCR5 (3A9, BD), CXCR5 (MU5UBEE, Invitrogen), PD-1 (EH12.2H7, BioLegend), ICOS (C398.4A, BioLegend), CCR7 (150503, BD), α4β7 (A4B7, NHP Reagent Resource), LAG3 (3DS223H, eBioscience), Tim-3 (F38-2E2, BioLegend), TIGIT (MBSA43, Invitrogen), CD20 (2H7, BioLegend), CD19 (J3-119, Beckman Coulter), HLA-DR (L243, BioLegend), CD10 (HI10A, BioLegend), CD21 (B-ly4, BD), CD27 (O323, BioLegend), IgD (IADB6, Southern Biotech), IgG (G18-145, BD), IL-21R (2G1-K12, BioLegend), Ki-67 (B56, BD), BCL6 (IG191E/A8, BioLegend), CD80 (2D10, BioLegend), CD56 (B159, BD), CD45 (D058-1283, BD), CD14 (MoP9, BD), CD16 (3G8, BD), CCR2 (48607, R&D Systems), CD11b (ICRF44, BioLegend), CD11c (3.9, Invitrogen), PD-L1 (29E.2A3, BioLegend), PD-L2 (24F.10C12, BioLegend), CD80 (2D10, BioLegend), CD86 (FUN-1, BD), CD141 (1A4, BD), CD163 (GHI/61, BioLegend), and CD123 (7G3, BD).

Kvistad D., Pallikkuth S., Sirupangi T., Pahwa R., Kizhner A., Petrovas C., Villinger F, & Pahwa S. (2021). IL-21 enhances influenza vaccine responses in aged macaques with suppressed SIV infection. JCI Insight, 6(20), e150888.

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