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Cd86 gl 1

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CD86 (GL-1) is a cell surface protein that functions as a co-stimulatory molecule. It is expressed on antigen-presenting cells and plays a role in the activation of T cells.

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Lab products found in correlation

Cd80 16 10a1, by BioLegend (6 mentions) Lsrfortessa, by BD (4 mentions) Cd45 30 f11, by BioLegend (4 mentions) Cd69 h1.2f3, by BioLegend (4 mentions) Cd3 145 2c11, by BioLegend (4 mentions) Mhcii m5 114, by BioLegend (3 mentions) Mp6 xt22, by BioLegend (3 mentions) Cd93 aa4, by BioLegend (3 mentions) Cd19 6d5, by BioLegend (3 mentions) Cd11c n418, by BioLegend (3 mentions) Cd11b m1 70, by BioLegend (3 mentions) Igd 11 26c 2a, by BioLegend (3 mentions) Tc11 18h10, by BD (2 mentions) Cd8a 53 6, by Thermo Fisher Scientific (2 mentions) Cd274 10f 9g2, by BioLegend (2 mentions) Xmg12, by BD (2 mentions) Cytofix cytoperm, by BD (2 mentions) Facscanto 2 cytometer, by BD (2 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Ionomycin, by Merck Group (2 mentions)

Spelling variants of this product

CD86 (clone GL-1, (9 citations) Anti-CD86 (clone GL-1, (7 citations) Anti-CD86 (GL-1, (7 citations) APC anti-mouse CD86 (clone GL-1, (5 citations) Anti-CD86-PE (clone GL-1, (4 citations) CD86 PE-Cy7 (clone GL-1; (3 citations) Anti mouse-CD86 (clone GL-1; (2 citations) Anti-mouse CD86 (GL-1, (2 citations) PE anti-mouse CD86 (clone GL-1, (2 citations) CD86-PerCP/Cy5.5 -clone GL-1- (2 citations)

16 protocols using cd86 gl 1

1

Multiparameter Flow Cytometry Protocol

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Flow cytometry data was acquired using the BD FACS Canto II (Becton-Dickinson) or BD LSR Fortessa (Becton-Dickinson). Data analysis was performed using Flowjo (Treestar Software). Antibodies were purchased from BD Biosciences, Affymetrix/eBioscience and Biolegend. Antibodies for CCR7 (4B12), CD11b (M1/70), CD11c (N418), CD44 (IM7), CD115 (AFS98), F4/80 (BM8), Gr-1 (RB6-8C5), Ly-6C (HK1.4), Ly-6G (1A8-Ly6G), MHC I (34-1-2S), OX40L (RM134L), PD-1 (J43), PD-L1 (MIH5), PD-L2 (TY25) were purchased from eBioscience. CD25 (PC61), CD40 (3/23), CD80 (16-10A1), CD103 (M290), and annexin V antibodies were from BD Biosciences. 7AAD was also from BD Biosciences. CD86 (GL-1) and MHC II (M5/114.15.2) antibodies were from Biolegend. gp34-tetramer was prepared by sequential addition of streptavidin APC or streptavidin-R-PE (Invitrogen) to biotinylated gp34 monomers (H-2Kb; AVYNFATC) generously provided by the NIH Tetramer Core Facility.
Tran C.W., Gold M.J., Garcia-Batres C., Tai K., Elford A.R., Himmel M.E., Elia A.J, & Ohashi P.S. (2020). Hypoxia-inducible factor 1 alpha limits dendritic cell stimulation of CD8 T cell immunity. PLoS ONE, 15(12), e0244366.
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2

Pancreatic Lymph Node Cell Profiling

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Dissociated cells from pancreatic lymph nodes were suspended in buffer containing 2% FBS in PBS. To stain for surface antigens, cells were incubated with specific antibodies to F4/80 (BM-8, Biolegend), CD11c (HL3, BD Pharmigen), CD4 (RM4–5, Biolegend), CD45 (30- F11, BD Biosciences), CD8a (53–6.7, eBioscience), CD80 (16-10A1, Biolegend), CD274 (10F.9G2, Biolegend), CD86 (GL-1, Biolegend), and MHC-II (M5/114.15.2, Biolegend) or the appropriate isotype controls for 30 min. For the intracellular staining, cells were stimulated with 100ng/mL PMA (Sigma Aldrich), 500ng/ml Ionomycin (Sigma Aldrich), and Golgi Stop Plug (BD Pharmigen, 1/1000). After stimulation, cells were first stained for surface antigens, then fixed and permeabilized with BD Cytofix/Cytoperm™ (BD Pharmigen), according to the manufacturer’s recommendations. Cells were then incubated with specific antibodies to TNF-α (MP6-XT22, Biolegend), IL-1β (NJTEN3, Thermo Fisher Scientific), IL-17 (TC11-18H10, BD Pharmigen), IFN-γ (XMG12, BD Pharmigen), and FoxP3 (MF23, BD Pharmigen). After staining, the cells were washed, filtered, and analyzed on a FACS Canto II cytometer (BD). Data were analyzed using FlowJo software (Tree Star).
Piñeros A.R., Kulkarni A., Gao H., Orr K.S., Glenn L., Huang F., Liu Y., Gannon M., Syed F., Wu W., Anderson C.M., Evans-Molina C., McDuffie M., Nadler J.L., Morris M.A., Mirmira R.G, & Tersey S.A. (2022). Proinflammatory signaling in islet β cells propagates invasion of pathogenic immune cells in autoimmune diabetes. Cell reports, 39(13), 111011.
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3

In vivo T cell Immunophenotyping

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In vivo labeling of T cells with fluorescent antibodies was performed as previously described (Turner et al., 2014). Briefly, mice were administered, PE- or Alexa 647-conjugated anti-CD90.2 (anti-Thy1.2; clone 53-2.1, BioLegend), or PE-conjugated anti-CD45.2 antibody (clone 104, Biolegend), intravenously, and 7–10 minutes later lungs were perfused, dissected and digested in RPMI1640 medium with collagenase D, DNAse and Trypsin inhibitor for 1 hour at 37°C. Mediastinal lymph nodes and spleen were dissected and manually disrupted to generate single cell suspensions. Cell suspensions were stained with fluorescent-conjugated antibodies for CD4 (clones RM4-5, BD Biosciences, and GK1.5, eBioscience), CD8 (clone 53-6.7, BioLegend), CD11a (clone M17/4, BioLegend),CD11b (M1/70 eBioscience), CD11c (N418, eBioscience), CD25 (PC61.5, eBioscience), CD45 (30-F11, BioLegend), CD69 (clone H1.2F3, eBioscience), CD86 (GL-1, Biolegend), CD103 (clone 2E7, eBioscience) and Anti-Mouse MHC Class II (I-A/I-E) (clone M5/114.15.2, eBioscience) was performed according to manufacturers’ protocol. Stained cells were analyzed using the BD LSRII flow cytometer and flow-jo software (Treestar, Ashland, OR). Absolute cell numbers were determined by flow cytometry using CountBright Absolute Counting Beads (Invitrogen) according to manufacturer’s protocol.
Turner D.L., Goldklang M., Cvetkovski F., Paik D., Trischler J., Barahona J., Cao M., Dave R., Tanna N., D’Armiento J.M, & Farber D.L. (2018). Biased generation and in situ activation of lung tissue-resident memory CD4 T cells in the pathogenesis of allergic asthma. Journal of immunology (Baltimore, Md. : 1950), 200(5), 1561-1569.
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4

Quantifying cDC Activation in Spleen

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Single cell suspensions of splenocytes were acquired with a MACSQuant (Miltenyi) flow cytometer and analyzed using FlowJo software (Tree Star). The following antibodies were used for quantifying cDCs and measuring cDC activation: TCRβ (H57-597) and CD11c (N418) from eBiosciences; CD19 (RA3-6B2), MHC II (M5/114.15.2), and CD86 (GL-1) from Biolegend. Zombie-NIR (Biolegend) was used for exclusion of dead cells. For evaluating cDC activation, splenocytes were processed 4 h following transfusion or i.v. injection of LPS.
Gibb D.R., Calabro S., Liu D., Tormey C.A., Spitalnik S.L., Zimring J.C., Hendrickson J.E., Hod E.A, & Eisenbarth S.C. (2016). The Nlrp3 Inflammasome Does Not Regulate Alloimmunization to Transfused Red Blood Cells in Mice. EBioMedicine, 9, 77-86.
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5

Immunophenotyping Mouse Immune Cells

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Mouse Abs and isotype control Abs (IgG1, IgG2a or IgG2b), CD11c (HL3), CD4 (GK1.5), CD8α (YTS169.4), CD40 (3/23), CD80 (16-10A1), and CD86 (GL-1) were obtained from BioLegend (Snd Diego, CA, USA); anti-MHC class I (AF6–88.5.3) and anti-MHC class II (M5/114.15.2) Abs were obtained from eBioscience (San Diego, CA, USA).
Xu L., Zhang W., Park H.B., Kwak M., Oh J., Lee P.C, & Jin J.O. (2019). Indocyanine green and poly I:C containing thermo-responsive liposomes used in immune-photothermal therapy prevent cancer growth and metastasis. Journal for Immunotherapy of Cancer, 7, 220.
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6

Multiparametric Flow Cytometry Analysis

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Cell suspensions from mouse liver, BM (one femur and tibia), spleen, thymus, mesenteric lymph nodes, and peritoneal cavity were processed as previously described (47 (link)), and data were acquired on a FACSCanto10c (BD) and analyzed with FlowJo Software (Tree Star).
The following anti-mouse antibodies were used: B220 (RA3-6B2, BioLegend), BAFFR (7H22-E16,BD), BP-1 (6C3, BioLegend), CD3e (145-2C11, eBioscience), CD4 (GK1.5, eBioscience), CD5 (53-7.3, BioLegend), CD8 (53-6.7, BioLegend), CD9 (MZ3, BioLegend), CD11b (M1/70, eBioscience), CD19 (6D5, 1D3, BioLegend), CD21/CD35 (7E9, BioLegend), CD23 (B3B4, BioLegend), CD24 (M1/69, BioLegend), CD25 (PC61, Biolgend), CD43 (S7, BioLegend), CD44 (IM7, BioLegend), CD45.1 (A20, BioLegend), CD45.2 (104, BioLegend), CD69 (H1.2F3, BioLegend), CD80 (16-10A1, BioLegend), CD86 (GL1, BioLegend), CD93 (AA4.1, BioLegend), DAPI (BIOTIUM), Gr-1 (RB6-8C5, eBioscience), IgDa (AMS-9.1, BioLegend), IgD (11-26c, BioLegend), IgMa (DS-1, MA-69, BioLegend), IgM (Il/41, RMM-1, BioLegend), TCRγδ (GL3, BioLegend), Live dead dye (Zombie Aqua Dye, BioLegend), Live dead dye (Zombie NIR, BioLegend), pERK (4B11B69, BD), pPLCγ2 (K86-1161, BD), Lin28b (AP1485C, ABGENT), and anti-rabbit IgG (BioLegend).
Xu X., Deobagkar-Lele M., Bull K.R., Crockford T.L., Mead A.J., Cribbs A.P., Sims D., Anzilotti C, & Cornall R.J. (2020). An ontogenetic switch drives the positive and negative selection of B cells. Proceedings of the National Academy of Sciences of the United States of America, 117(7), 3718-3727.
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7

Flow Cytometry Immune Cell Profiling

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Single-cell suspensions were stained
with the fixable viability dye eFluor 780 (eBioscience) for 30 min
at 4 °C, and Fc receptors were blocked with an anti-CD16/32 (BioLegend)
blocking antibody prior to surface staining with monoclonal antibodies.
Antibodies used for surface staining are as listed: mouse: CD45 (30-F11,
BioLegend), lineage (Lin) markers [CD3 (145-2C11, BioLegend), CD19
(6D5, BioLegend), CD11b (M1/70, BioLegend), CD11c (N418, BioLegend)],
and CD86 (GL-1, BioLegend). For CFSE staining, cells were stained
with 5 μM CFSE according to the manufacturer’s protocol.
Data were acquired on LSR II (BD Biosciences) and analyzed with FlowJo
v. 10.1 software (TreeStar).
González-Cuesta M., Lai A.C., Chi P.Y., Hsu I.L., Liu N.T., Wu K.C., García Fernández J.M., Chang Y.J, & Ortiz Mellet C. (2023). Serine-/Cysteine-Based sp2-Iminoglycolipids as Novel TLR4 Agonists: Evaluation of Their Adjuvancy and Immunotherapeutic Properties in a Murine Model of Asthma. Journal of Medicinal Chemistry, 66(7), 4768-4783.
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8

Pancreatic Lymph Node Cell Profiling

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Dissociated cells from pancreatic lymph nodes were suspended in buffer containing 2% FBS in PBS. To stain for surface antigens, cells were incubated with specific antibodies to F4/80 (BM-8, Biolegend), CD11c (HL3, BD Pharmigen), CD4 (RM4–5, Biolegend), CD45 (30- F11, BD Biosciences), CD8a (53–6.7, eBioscience), CD80 (16-10A1, Biolegend), CD274 (10F.9G2, Biolegend), CD86 (GL-1, Biolegend), and MHC-II (M5/114.15.2, Biolegend) or the appropriate isotype controls for 30 min. For the intracellular staining, cells were stimulated with 100ng/mL PMA (Sigma Aldrich), 500ng/ml Ionomycin (Sigma Aldrich), and Golgi Stop Plug (BD Pharmigen, 1/1000). After stimulation, cells were first stained for surface antigens, then fixed and permeabilized with BD Cytofix/Cytoperm™ (BD Pharmigen), according to the manufacturer’s recommendations. Cells were then incubated with specific antibodies to TNF-α (MP6-XT22, Biolegend), IL-1β (NJTEN3, Thermo Fisher Scientific), IL-17 (TC11-18H10, BD Pharmigen), IFN-γ (XMG12, BD Pharmigen), and FoxP3 (MF23, BD Pharmigen). After staining, the cells were washed, filtered, and analyzed on a FACS Canto II cytometer (BD). Data were analyzed using FlowJo software (Tree Star).
Piñeros A.R., Kulkarni A., Gao H., Orr K.S., Glenn L., Huang F., Liu Y., Gannon M., Syed F., Wu W., Anderson C.M., Evans-Molina C., McDuffie M., Nadler J.L., Morris M.A., Mirmira R.G, & Tersey S.A. (2022). Proinflammatory signaling in islet β cells propagates invasion of pathogenic immune cells in autoimmune diabetes. Cell reports, 39(13), 111011.
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9

Analyzing Virus-Specific T Cell Responses

Cited 2 times
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After animal sacrifice and perfusion with ice cold PBS, half of the brain of mice was collected and homogenized at defined times p.i. and leukocytes enriched on a Percoll gradient as previously described (Blanc et al., 2014 (link)). Cells were stained using the following antibodies: CD4 (GK1.5, Biolegend), CD3 (145–2 C11, Biolegend), IFN-γ (XMG1.2, Biolegend), FoxP3 (FJK-16s, eBioscience), IL-10 (JES5-16E3, Biolegend), CD8 (53-6.7, Biolegend), CD45 (30-F11, Biolegend), F4/80 (BM8, Biolegend), CD11b (M1/70, Biolegend), CD40 (HM40-3, Biolegend), CD86 (GL-1, Biolegend), MHCI (M1/42, Biolegend), and MHCII (M5/114.15.2, Biolegend). Virus-specific T cells were determined via flow cytometric analysis through either intracellular staining for IFN-γ or defined tetramers specific for immunodominant CD4 +and CD8+T cell-specific viral epitopes (Chen et al., 2014 (link); Stiles et al., 2006 (link)). Samples were analyzed using BD LSRFortessa and FACSDiva software and data was measured using FlowJo. 
Brown D.G., Soto R., Yandamuri S., Stone C., Dickey L., Gomes-Neto J.C., Pastuzyn E.D., Bell R., Petersen C., Buhrke K., Fujinami R.S., O'Connell R.M., Stephens W.Z., Shepherd J.D., Lane T.E, & Round J.L. (2019). The microbiota protects from viral-induced neurologic damage through microglia-intrinsic TLR signaling. eLife, 8, e47117.
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10

Surface Marker Analysis of Immune Cells

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For surface markers analysis, live cells were re-suspended in 1 × PBS and stained with anti-mouse CD45 (30-F11, Biolegend), CD11b (M1/70, R&D Systems), F4/80 (BM8, Biolegend), CD86 (GL-1, Biolegend), MHC-II (M5/114.15.2, Biolegend), CD117 (2B8, Biolegend), CD115 (AFS98, Biolegend), CD16/32 (93, Biolegend), CD34 (HM34, Biolegend), Sca1 (D7, Biolegend), SIRPα (P84, biolegend), Lin (Stem Cell), and APC-Cy7 Streptavidin (Biolegend) at 4°C for 30 min. The concentration at each antibody was used as the product protocol recommended. For intracellular cytokine staining, cells were fixed and permeabilized with Fixation and Permeabilization Kit (eBioscience) at room temperature for 40 min and labeled with anti-mouse CD206 (C068C2, Biolegend). Multicolor FACS analysis was performed on an LSR Fortessa Analyzer (BD Biosciences). All data analysis was performed using the flow cytometry analysis program FlowJo-V10.
Zhao C.C., Han Q.J., Ying H.Y., Gu X.X., Yang N., Li L.Y, & Zhang Q.Z. (2022). TNFSF15 facilitates differentiation and polarization of macrophages toward M1 phenotype to inhibit tumor growth. Oncoimmunology, 11(1), 2032918.
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