Mouse anti-human CD11c-PE/Cy7, EPCAM-PE/CF594, CD86-BV510, HLA-DR PerCP/Cy5.5, PD-L1-BV421, PD-L2-APC, and CD83-APC were purchased from Biolegend (San Diego, CA). Fixable Viability Dye eFluor™ 780 was purchase from eBioscience (San Diego, CA) and mouse anti-human CD56-FITC were purchased from BD Biosciences (San Jose, CA). Recombinant human IL-4 and GM-CSF were purchased from R&D systems (Minneapolis, MN) reconstituted according manufacturer instructions and kept at −80 Celsius freezer in aliquots. 2′, 3′- cGAMP sodium salt was purchased from Tocris (Bristol, UK). Cetuximab (anti-EGFR mAb, IgG1) was kindly provided by Bristol-Meyers Squibb and was used at a concentration of 10 ug/mL in all our experiments. Human IgG1 Kappa-LE/AF was purchased from Sourthern Biotech (Birmingham, AL). Western blot antibodies included rabbit anti-human STING, pIRF3, and total IRF3 were purchased from Cell Signaling (Danvers, MA). Mouse anti-human β-actin was purchased from Bio Rad (Hercules, CA). Secondary goat anti-rabbit and anti-mouse antibodies were purchased from LI-COR biosciences (Lincoln, NE). STING primary IHC antibody was purchased from Abcam (Cambridge, MA). Human IFN-gamma Quantikine ELISA Kit was purchased from R&D systems (Minneapolis, MN).
Cd86 bv510
Manufactured by BioLegend
CD86-BV510 is a fluorochrome-conjugated antibody that binds to the CD86 protein, also known as B7-2. CD86 is a co-stimulatory molecule expressed on the surface of antigen-presenting cells, such as dendritic cells, macrophages, and B cells. The BV510 fluorochrome is used to label the antibody, enabling its detection in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Human ifn gamma quantikine elisa kit, by R&D Systems (2 mentions)
P irf3, by Cell Signaling Technology (2 mentions)
Pd l1 bv421, by BioLegend (2 mentions)
Total irf3, by Cell Signaling Technology (2 mentions)
Mouse anti human cd11c pe cy7, by BioLegend (2 mentions)
Epcam pe cf594, by BioLegend (2 mentions)
Pd l2 apc, by BioLegend (2 mentions)
Mouse anti human cd56 fitc, by BD (2 mentions)
Rabbit anti human sting, by Cell Signaling Technology (2 mentions)
Mouse anti human β actin, by Bio-Rad (2 mentions)
Sting primary ihc antibody, by Abcam (2 mentions)
Recombinant human il 4, by R&D Systems (2 mentions)
Cd83 apc, by BioLegend (2 mentions)
Fixable viability dye efluor 780, by Thermo Fisher Scientific (2 mentions)
Hla dr percp cy5, by BioLegend (2 mentions)
Gm csf, by R&D Systems (2 mentions)
Hla dr alexa fluor 488, by BioLegend (1 mentions)
Pd l1 pe, by BioLegend (1 mentions)
Cyan adp flow cytometer, by Beckman Coulter (1 mentions)
Cd80 apc, by BioLegend (1 mentions)
4 protocols using cd86 bv510
1
Multiparameter Flow Cytometry Panel
2
Multiparameter Flow Cytometry Panel
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Mouse anti-human CD11c-PE/Cy7, EPCAM-PE/CF594, CD86-BV510, HLA-DR PerCP/Cy5.5, PD-L1-BV421, PD-L2-APC, and CD83-APC were purchased from Biolegend (San Diego, CA). Fixable Viability Dye eFluor™ 780 was purchase from eBioscience (San Diego, CA) and mouse anti-human CD56-FITC were purchased from BD Biosciences (San Jose, CA). Recombinant human IL-4 and GM-CSF were purchased from R&D systems (Minneapolis, MN) reconstituted according manufacturer instructions and kept at −80 Celsius freezer in aliquots. 2′, 3′- cGAMP sodium salt was purchased from Tocris (Bristol, UK). Cetuximab (anti-EGFR mAb, IgG1) was kindly provided by Bristol-Meyers Squibb and was used at a concentration of 10 ug/mL in all our experiments. Human IgG1 Kappa-LE/AF was purchased from Sourthern Biotech (Birmingham, AL). Western blot antibodies included rabbit anti-human STING, pIRF3, and total IRF3 were purchased from Cell Signaling (Danvers, MA). Mouse anti-human β-actin was purchased from Bio Rad (Hercules, CA). Secondary goat anti-rabbit and anti-mouse antibodies were purchased from LI-COR biosciences (Lincoln, NE). STING primary IHC antibody was purchased from Abcam (Cambridge, MA). Human IFN-gamma Quantikine ELISA Kit was purchased from R&D systems (Minneapolis, MN).
3
Flow Cytometry Analysis of Dendritic Cells
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iDCs and mDCs harvested at day 8 were analysed for phenotypic markers by flow cytometry. Cells were fixed using Human FoxP3 Buffer Set (BD Biosciences, 560098), and stained with anti-human: CD80 APC, CD86 BV510, HLA-DR AlexaFluor488, PD-L1 PE, (all BioLegend, 305220, 305431, 307656, 329705, respectively). Cells were then analysed using a cyAn ADP flow cytometer (Beckman Coulter).
4
Characterizing Macrophage Surface Markers
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To examine surface marker expression, BMDMs were detached using citrate buffer (17 mM tri-Sodium citrate dehydrate and 135 mM potassium chloride in water), transferred to V-bottom well plates, incubated with 1:50 anti-CD16/CD32 Fc-block and stained for 20 min at room temperature in the dark with antibodies. Antibodies were directed to CD40-APC (Biolegend, 1:100), MHC-II-PerCP/Cy5.5 (Biolegend, 1:200), CD80-BV650 (Biolegend, 1:100) and CD86-BV510 (Biolegend, 1:100) for LPS-stimulated macrophages or directed to CD71-PE (BD PharMingen, 1:250), CD206-APC (Biolegend, 1:250), CD273-PE (BD PharMingen, 1:250), CD301-AF647 (Serotec, 1:250) or isotype controls (Biolegend) for IL-4 stimulated macrophages (IGG2A-PE, IGG2A-APC), diluted in staining buffer (PBS with 0.5% BSA and 2.5 mM ethylenediaminetetraacetic acid; EDTA). After staining, cells were washed and resuspended in staining buffer, measured on a Beckman Coulter CytoFLEX or BD LSR Fortessa and analyzed using FlowJo (Tree Star). Surface expression was calculated as ΔMFI = (median fluorescence intensity)positive staining – (median fluorescence intensity)isotype or FMO control. MFI controls (isotype or FMO) were measured on a pool of all samples.
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