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Anti cd86

Manufactured by ABclonal
Sourced in United States

Anti-CD86 is a laboratory reagent used for the detection and analysis of CD86, a protein expressed on the surface of activated antigen-presenting cells. It can be used in various immunological studies and flow cytometry applications.

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3 protocols using anti cd86

1

Immunofluorescence Staining of Lung and THP-1 Cells

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Immunofluorescence staining was performed as previously described with minor modification [28 (link)] [PMID: 33,472,663]. The lung tissue or THP-1 cell sections were blocked with 1% bovine serum albumin (BSA) (Sangon Biotech, Shanghai, China) for 15 min and incubated with anti-CD68 (diluted 1:50; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-CD86 (diluted 1:100; ABclonal Technology, Wuhan, China)), anti-iNOS (diluted 1:100; Affinity Biosciences, Changzhou, China), anti-NLRP3 (diluted 1:100; ABclonal Technology, Wuhan, China), anti-ASC (diluted 1:50; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and anti-p65 (diluted 1:400; CST, Danvers, MA, USA) overnight. Subsequently, the slices were incubated with corresponding secondary antibodies (Cy3: goat-anti rabbit, diluted 1:200, Invitrogen, Carlsbad, CA, USA; FITC: goat-anti-mouse, diluted 1:200, Abcam, Cambridge, UK), followed by counterstaining with 4’, 6-diamidino-2-phenylindole (DAPI) (Aladdin, Shanghai, China). Fluorescence microscope (Olympus, Tokyo, Japan) was used to capture stained images.
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2

Immunohistochemical Analysis of Tumor Markers

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For immunohistochemistry (IHC), 5-µm paraffin-embedded sections of patient/mice tumors were baked at 60°C for 1 h, deparaffinized in xylene, and rehydrated in a graded series of ethanol solutions. Antigens were unmasked by microwave heating of the samples in 10 mM sodium citrate buffer (pH 6.0) for 15 m (5 m each for 3 times), and the reaction was quenched using hydrogen peroxide (3%). After washing with phosphate-buffered saline, samples were incubated with the following primary antibodies, respectively, at 4°C overnight: anti-FCER1G (A12889; ABclonal, Woburn, MA, USA), anti-CD86 (A1199; ABclonal), anti-CyclinB1 (A19037; ABclonal), anti-TPA (10147-1-AP; Proteintech, Rosemont, IL, USA), anti-ITGA5 (27224-1-AP; Proteintech), or anti-CD40 (66965-1-Ig; Proteintech). DAB (3,3′-diaminobenzidine) was used as a detection system.
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3

Quantification of CD86 Expression on RAW264.7 and THP-1 Cells

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We seeded RAW264.7 cells on 24-well plates (1 × 105 cells/well) and used BSHXD and LPS to process for a certain time. Then we fixed the cells in the pore plate and incubated overnight at 4 °C with anti-CD86 (1:1000, A1199, Abclonal, Wuhan, China) antibody. The Goat Anti-Mouse IgG H&L (Alexa Fluor® 555) (1:1000, ab150114, Abcam) were incubated with cell for 1 h at 37 °C.100 μl Alexa Fluor® 488 Phalloidin (1:20, #8878s, CST, Danvers, USA) added to the cells. The nuclei were stained with DAPI. Fluorescence images were obtained using immunofluorescence microscopy (Thermo Fisher Scientific, Bothell, WA, USA). The THP-1 cells were immunofluorescence stained with the same procedure.
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