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Anti cd86 gl 1

Manufactured by BioLegend
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Anti-CD86 (GL-1) is a lab equipment product that functions as an antibody specific for the CD86 cell surface protein. CD86 is a co-stimulatory molecule expressed on antigen-presenting cells and plays a role in T cell activation.

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Lab products found in correlation

Anti cd43 1b11, by BioLegend (3 mentions) Anti cd11b m1 70, by BioLegend (3 mentions) Foxp3 fixation and permeabilization kit, by Thermo Fisher Scientific (2 mentions) Anti f4 80 bm8, by BioLegend (2 mentions) Anti cd28 37, by BioLegend (2 mentions) Anti cd80 16 10a1, by BioLegend (2 mentions) Facscanto 2 flow cytometer, by BD (2 mentions) Anti cd19 1d3, by BioLegend (1 mentions) Ki67 sola15, by BD (1 mentions) Anti cd138 281.2, by BD (1 mentions) Cd62l mel 14, by BioLegend (1 mentions) Anti cd4 rm4 5, by BioLegend (1 mentions) Anti cd44 im7, by BD (1 mentions) Anti cd11c hl3, by BD (1 mentions) Anti nk1.1 pk136, by BioLegend (1 mentions) Anti cd45 30 f11, by BD (1 mentions) Anti cd25 pc61, by Thermo Fisher Scientific (1 mentions) Anti cd4 rm4 5, by Thermo Fisher Scientific (1 mentions) Anti ctla 4 uc10 4b9, by BioLegend (1 mentions) Anti cd8a 53 6, by BioLegend (1 mentions)

7 protocols using anti cd86 gl 1

1

Multiparameter Flow Cytometry Analysis

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The following antibodies were used for cell surface staining: anti-CD4 (RM4-5, BioLegend), anti-CD19 (1D3, BioLegend), anti-CD38 (90, eBio), anti-CD138 (281–2, BD), anti-CD86 (Gl1, Biolegend), anti-Ter119 (TER-119, eBio), anti-GL7 (GL-7, eBio), anti-mouse IgD (11–26, eBio) anti-mouse IgM (11/41, eBio), anti-PD1 (J43, eBio), anti-CD43 (1B11, BioLegend), CD62L (MEL-14, BioLegend). Following surface staining, intracellular staining was performed with the FOXP3 permeabilization and fixation kit from eBiosciences according to the manufacturers recommendations using the following antibodies: Ki-67 (SolA15, BD) or FOXP3 (FJK-16s, eBio). FOXP3 expression in cells from FOXP3-DTR mice was also analyzed by GFP expression. Lymphocyte populations were initially gated on the basis of forward scatter versus side scatter. B cells were gated as CD19+CD4- cells within the lymphocyte gate. GC B cells were gated as CD38-GL7+ cells within the B cell gate and plasmablasts were gated as CD38+CD138+ cells within the B cell gate. CD4 T cells were gated as CD4+ cells within the lymphocyte gate. See S1 Fig for gating strategies. Flow cytometic data were collected with an LSRII (BD Biosciences) and analyzed using FlowJo software (Tree Star).
Moore T.C., Messer R.J, & Hasenkrug K.J. (2018). Regulatory T cells suppress virus-specific antibody responses to Friend retrovirus infection. PLoS ONE, 13(4), e0195402.
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2

Multiparametric Flow Cytometry Analysis

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The harvested spleens were mashed and single-cell suspensions of the spleens were stained with the following surface antibodies: anti-CD45 (30-F11, BD Biosciences), LIVE/DEAD Fixable Blue cell stain (Invitrogen), anti-CD19 (ID3, BD Biosciences), anti-CD8a (53-6.7, BioLegend), anti-CD27 (LG7F9, eBioscience), anti-CD11b (M1/70, BioLegend), anti-CD11c (HL3, BD Biosciences), anti-CD86 (GL1, BioLegend), anti-NK1.1 (PK136, BioLegend), anti-NKp46 (29A1.4, BioLegend), anti-MHC Class II (M5/114.15.2, BD Biosciences), anti-F4/80 (BM8, BioLegend), anti-CD80 (16-10A1), anti-CD3 (17A2, BioLegend), anti-CD4 (RM4-5, Invitrogen), anti-CTLA-4 (UC10-4B9, BioLegend), anti-PD-1 (29F.1A12, eBioscience), anti-CD28 (37.51, BioLegend), anti-CD44 (IM7, BD Biosciences), anti-CD43 (1B11, BioLegend), anti-CD47 (miap301, BioLegend), anti-CD62L (MEL-14, BioLegend), anti-CD25 (PC61.5, eBioscience), and anti-CD107a (1D4B, BioLegend). Intracellular staining was then performed with a FOXP3 permeabilization and fixation kit (eBioscience) according to manufacturer’s recommendations using the following antibodies: Ki-67 (B56; BD Biosciences) and GrB (QA16A02, BioLegend). Flow cytometric data were collected with a Beckman Coulter Cytoflex LX (6-L NUV) flow cytometer and analyzed using FlowJo software (Tree Star).
Rao D., O’Donnell K.L., Carmody A., Weissman I.L., Hasenkrug K.J, & Marzi A. (2021). CD47 expression attenuates Ebola virus-induced immunopathology in mice. Antiviral research, 197, 105226.
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3

Multiparametric Immune Cell Analysis

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Cell surface and intracellular staining of CD8+ T cells was performed as previously described (41 (link), 42 (link)) using the following antibodies (BioLegend, San Diego, CA, USA): anti-CD8 (53-6.7), anti-GzmB (GB11), anti-IFNγ (XMG1.2), anti-IL-2 (JES6-5H4) and anti-TNFα (MP6-XT22). Dead cells were excluded from analysis (positive for fixable viability dye, Thermo Scientific). For phenotypic analysis of BM-DCs, surface staining was performed with anti-CD11b (M1/70, BioLegend), anti-CD11c (N418, Miltenyi Biotec), anti-CD80 (16-10A1, BioLegend), anti-CD86 (GL-1, BioLegend) and anti-MHC class II (M5/114.15.2, Miltenyi Biotec), and intracellular staining was performed using anti-IL-6 (MP5-20F3, BioLegend). Fluorescence minus one (FMO) controls were used for all conditions. Data were acquired on a FACS Canto II flow cytometer (BD Biosciences, Heidelberg, Germany) and analyses were performed using Flow Jo (BD Biosciences) software.
Dickow J., Francois S., Kaiserling R.L., Malyshkina A., Drexler I., Westendorf A.M., Lang K.S., Santiago M.L., Dittmer U, & Sutter K. (2019). Diverse Immunomodulatory Effects of Individual IFNα Subtypes on Virus-Specific CD8+ T Cell Responses. Frontiers in Immunology, 10, 2255.
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4

Synthesis and Characterization of Pam2LPs

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Pam2LPs were synthesised, as described previously.14 (link) Poly(I:C) was purchased from GE Healthcare Life Sciences (27-4732-01).
EndoGrade® Ovalbumin was purchased from Hyglos (321001). Anti-TLR2 Ab (Clone:
6C2; cat. no. 12-19021-80) and a mouse IFN-γ ELISA KIT (88-7314) were purchased
from eBioscience. ViaProbe was purchased from BD Biosciences (555816).
Carboxyfluorescein diacetate succinimidyl ester (CFSE) was purchased from
Molecular Probes (C1157). The following Abs were purchased from BioLegend:
anti-CD3ɛ (145-2C11, 100314), anti-CD8α (53-6.7, 100712), anti-CD11c (N418,
117310), anti-CD16/32 (93, 101302), anti-CD28 (37.51, 102111), anti-CD40 (3/23,
124607), anti-CD80 (16-10A1, 104707), anti-CD86 (GL-1, 105005) and anti-TCR
Vβ5.1,5.2 (MR9-4, 139504). An anti-TLR6 Ab (418601, MAB1533) was purchased from
R&D Systems.
Takeda Y., Azuma M., Hatsugai R., Fujimoto Y., Hashimoto M., Fukase K., Matsumoto M, & Seya T. (2018). The second and third amino acids of Pam2 lipopeptides are key for the proliferation of cytotoxic T cells. Innate Immunity, 24(5), 323-331.
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5

Multicolor Flow Cytometry Profiling

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Cell suspensions were stained on ice for 20 min in the dark with various combinations of fluorochrome-conjugated antibodies, including anti-CD45 (30-F11), anti-CD11b (M1/70), anti-F4/80 (BM8), anti-Ly6G (1A8), anti-Ly6C (HK1.4), anti-MHC II (M5/114.15.2), anti-CD11c (N418), anti-CD206 (MMR), and anti-CD86 (GL-1) (all from BioLegend, San Diego, CA, USA). Samples were acquired on a FACS Canto II flow cytometer (BD, Franklin Lakes, NJ, USA). Data were analyzed using the FlowJo™ Software (BD).
Hong S.M., Lee A.Y., Hwang S.M., Ha Y.J., Kim M.J., Min S., Hwang W., Yoon G., Kwon S.M., Woo H.G., Kim H.H., Jeong W.I., Shen H.M., Im S.H., Lee D, & Kim Y.S. (2022). NAMPT mitigates colitis severity by supporting redox-sensitive activation of phagocytosis in inflammatory macrophages. Redox Biology, 50, 102237.
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6

Flow Cytometric Analysis of Activated Immune Cells

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Cell surface staining was performed using the following antibodies: anti-CD11b (M1/70, Miltenyi Biotec), anti-CD11c (N418, BioLegend) anti-CD3 (17A2, eBioscience), anti-CD49b (DX5, eBioscience), anti-CD69 (H1.2 F3, eBioscience), anti-CD86 (GL-1, BioLegend), anti-MHC-II (M5/114.15.2, Miltenyi Biotec), anti-NK1.1 (PK136, BD Bioscience). Dead cells were excluded from analysis (positive for fixable viability dye, eBioscience). For detection of activated T cells splenocytes were stained with anti-CD4 (GK1.5, eBioscience), anti-CD8 (53–6.7, eBioscience), anti-CD43 (1B11, BioLegend) and anti-CD62L (MEL-14, eBioscience). Intracellular IFN-γ (XMG1.2, Miltenyi Biotec), IL-2 (JES6-5H4, Miltenyi Biotec) and TNF-α (MP6-XT22, BioLegend) staining was performed as described [28 (link),29 (link)]. Data were acquired on LSR II flow cytometer (BD Bioscience) and analyses were performed using FACSDiva (BD Bioscience) and Flow Jo (Tree Star, USA) software.
Gibbert K., Francois S., Sigmund A.M., Harper M.S., Barrett B.S., Kirchning C.J., Lu M., Santiago M.L, & Dittmer U. (2014). Friend retrovirus drives cytotoxic effectors through Toll-like receptor 3. Retrovirology, 11, 126.
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7

Multiparametric Flow Cytometry Analysis

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Cellular suspensions from the tissues were prepared as described previously in 2. The following antibodies were used: anti-CD19 (1D3, #45-0193-80, eBioscience), anti-B220 (RA3-6B2, #RM2630, Life Technologies), anti-CD45 (104, #109825, Biolegend), anti-CD1d (1B1, #123507, Biolegend), anti-CD140 (APA5, #135905, Biolegend), anti-CD21 (7E9, #123419, Biolegend), anti-CD5 (53-7.3, #100607, Biolegend), anti-AA4.1 (#17-5892, eBioscience), anti-CD138 (281-2, #142505, Biolegend), anti-CD206 (C068C2, Biolegend), anti-CD86 (GL-1, Biolegend), anti-F4-80 (BM8, Biolegend), anti-CD11b (M1-70, Biolegend). Dead cells were excluded by staining with Propidium Iodide (Sigma-Aldrich) or Aqua Live/Dead stain. Flow cytometry was performed on FACScalibur and LSRII II (BD Biosciences) instruments at NYU School of Medicine Flow Cytometry Core Facility and data was analyzed using FlowJo software.
Pylayeva-Gupta Y., Das S., Handler J.S., Hajdu C.H., Coffre M., Koralov S.B, & Bar-Sagi D. (2015). IL-35 producing B cells promote the development of pancreatic neoplasia. Cancer discovery, 6(3), 247-255.
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