For flow cytometry based quantification in Figure S1 , PD-L1 and CD80 were stained by PE anti-PD-L1 (eBioscience, 14-5983-82) and PE anti-CD80 (Biolegend, 305208) and their expression levels were quantified using the QUANTUM R-PE MESF kit (Bangs Laboratories Inc, 827), following manufacturer’s instructions. For immunoblot-based quantifications, total cell lysates were subjected to SDS-PAGE, transfected to a nitrocellulose membrane. Afterward, PD-L1 was probed by PE anti-PD-L1 (eBioscience, 14–598382) and detected by a Typhoon 5 Biomolecular Imager; CD80 was sequentially probed by anti-CD80 (Novus Biologicals, NBP2–25255) and DyLight488 anti-mouse IgG (Biolegend, 405310), then detected by Typhoon 5 Biomolecular Imager. Molecular densities were calculated assuming the following diameters:13 μm for Raji B cells (Hui et al., 2017 (link)) and 12.5 μm for DCs (Dumortier et al., 2005 (link)).
Anti cd80
Manufactured by Novus Biologicals
Anti-CD80 is a laboratory reagent used for research purposes. It is an antibody that binds to the CD80 protein, also known as B7-1, which is expressed on the surface of antigen-presenting cells. Anti-CD80 can be used in various experimental techniques to study the role of CD80 in cellular processes.
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Lab products found in correlation
Anti pd l1 pe, by Thermo Fisher Scientific (1 mentions)
Anti cd80 pe, by BioLegend (1 mentions)
Anti cd3 antibody, by Abcam (1 mentions)
Anti cd31, by Novus Biologicals (1 mentions)
Anti cd8 antibody, by Novus Biologicals (1 mentions)
Anti f4 80, by R&D Systems (1 mentions)
Anti cd206, by R&D Systems (1 mentions)
Anti cd4 antibody, by Abcam (1 mentions)
2 protocols using anti cd80
1
Quantification of PD-L1 and CD80 Levels
2
Immunohistochemical Analysis of Tumor Microenvironment
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Immediately after mice were sacrificed, tumor tissue was fixed in 10% formalin before paraffin embedding. Standard procedures were used for hematoxylin and eosin (H&E) staining. For immunostaining, formalin-fixed, paraffin-embedded tissue was treated with xylene, rehydrated with ethanol, and heated in a microwave with citric buffer to retrieve antigens. For blocking purpose, the tissues were incubated for 30 minutes, with 5% bovine serum albumin buffer. Followed by overnight incubation at 4° C, with primary antibodies: anti-CD3 antibody, anti-CD4 antibody, and anti-NKp46 antibody (Abcam, Cambridge, United Kingdom) at 1:100 dilutions, anti-CD8 antibody (Novus Biologicals, Minneapolis, MN) at 1:20 dilution, anti-CD31 (Novus Biologicals, Minneapolis, MN) at 1:100, anti-F4/80 (R&D system Minneapolis, MN) at 1:100, anti-CD206 (R&D system Minneapolis, MN) at 1:100, as well as anti-CD80 (Novus Biologicals, Minneapolis, MN) at 1:100. After washing, tissues were incubated with fluorescence-conjugated secondary antibodies at room temperature for 1 hr. Slides were prepared with antifade mountant with 4’,6-diamidino-2-phenylindole (DAPI).
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