Cd86 clone gl1

Manufactured by Thermo Fisher Scientific

CD86 (clone GL1) is a monoclonal antibody that binds to the CD86 antigen, also known as B7-2. CD86 is a costimulatory molecule expressed on antigen-presenting cells and plays a critical role in T-cell activation. This product is intended for research use only.

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Close spelling variants of this product

Anti-CD86-APC (clone GL1; CD86-FITC (clone GL1; FITC-anti-CD86 clone GL1 Anti-CD86 (clone GL1, Anti-CD86-PE-Cy5 (clone GL1, CD86 (GL1,

2 protocols using cd86 clone gl1

Characterization of Tumor-Infiltrating Lymphocytes

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Single cell suspensions from spleens and lymph nodes were prepared as described.6 (link) TIL were isolated from pooled tumors as described.26 (link) All flow cytometry experiments were performed at least 3 times. Single cell suspensions were incubated with mouse Fc receptor binding inhibitor for 10 minutes before staining with antibodies to CD45 (clone 30-F11), CD3 (clone 145-2C11), CD4 (clone GK1.5), CD8 (clone 53-6.7), CD19 (clone eBio1D3), CD86 (clone GL1), CD11b (clone M1/70), Gr-1 (clone RB6-8C5), CD69 (clone H1.2F3), CD44 (clone IM9), CD62L (clone MEL-14), and CD11c (clone N418; all from eBioscience, San Diego, CA) for 30 minutes. Flow cytometry was performed using FACS Calibur (BD Biosciences) and the lymphocyte population was selected by gating CD45⁺ cells. The data were analyzed using Flow Jo software (Tree Star, Ashland, OR). For tetramer staining, 10 μL of PE labeled HLA-A*02:01 Human HPV16 E7 tetramer (NIH Tetramer Core Facility) was added to 200 μL mouse lymphocyte suspension (1×106 cells per tube). After incubation for 30 minutes, cells were centrifuged and resuspended in phosphate-buffered saline with 1% paraformaldehyde and then analyzed by flow cytometry. PE labeled HLA-A*02:01 human mesothelin tetramer (NIH Tetramer Core Facility) was used as control.

Dai M., Hellstrom I., Yip Y.Y., Sjögren H.O, & Hellstrom K.E. (2018). Tumor Regression and Cure Depends on Sustained Th1 Responses. Journal of Immunotherapy (Hagerstown, Md. : 1997), 41(8), 369-378.

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Quantifying Dendritic Cell Maturation

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BMDC were generated from C57BL/6 mice as described above. 1x10⁶ DC were plated in each well of 6-well tissue culture-treated plates in a volume of 800ul. DC were transduced with Ad5-LMP1 or Ad5-GFP control (MOI = 50). DC were incubated with virus at 4°C for 1 h, followed by 3 h at 37°C. Complete media was then added to 2ml with a final concentration of 1ug/ml doxycyline. As a positive control, cytokine mix Mimic (5 ng/ml TNF-α (Peprotech), 5ng/ml IL-1b (Peprotech), 750ng/ml IL-6 (Peprotech), and 1ug/ml PGE2 (Sigma)) was used to mature DC. Cells were incubated for 36 hours at 37°C, harvested, and stained with the following antibodies: anti-mouse CD80 clone 16-10A1, CD86 clone GL1, CD40 clone 1C10, CD83 clone Michel-17, MHC Class II (I-A/I-E) clone M5-114.15.2, and CCR7 clone 4B12 (all from eBioscience). Tubes were also stained with hamster anti-mouse CD11c clone N418 PE-Cyanine7 conjugate (eBioscience) to allow for gating on CD11c+ DC. After flow cytometry analysis, the mean fluorescence intensity (MFI) for each antibody was calculated for CD11c+ dendritic cells under each experimental condition. FlowJo 7.6.4 flow cytometry analysis software (FlowJo, Ashland, OR) was used for analysis. Three independent wells were analyzed for each condition.

Termini J.M., Gupta S., Raffa F.N., Guirado E., Fischl M.A., Niu L., Kanagavelu S, & Stone G.W. (2017). Epstein Barr virus Latent Membrane Protein-1 enhances dendritic cell therapy lymph node migration, activation, and IL-12 secretion. PLoS ONE, 12(9), e0184915.

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