Following Fc block (eBiosciences, 93) cells were stained with the following antibodies: CD11a-PE (BioLegend, 2D7), CD18-FITC (BD Biosciences, C71/16) CD11c-PE-Cy7 (eBioscience, N418), CD11b-APC (BioLegend, M1/70), CD80-APC (eBioscience, 16-10A1), CCR7-PE (BioLegend, 4B12), CD40-PE (BioLegend, 3/23), CD86-FITC (BD Biosciences, GL1). FITC-conjugated antibodies were used at 1:100 dilution, all other antibodies were used at 1:200 dilution. Data were acquired using a LSRForetessa (BD) and analyzed using FlowJo software (TreeStar, USA).
Apc cd80
Manufactured by Thermo Fisher Scientific
Sourced in United States
The APC-CD80 is a flow cytometry reagent that binds to the CD80 antigen. It is used for the identification and quantification of CD80-expressing cells in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cd86 pe, by Thermo Fisher Scientific (6 mentions)
Cd86 apc, by Thermo Fisher Scientific (4 mentions)
Cd40 apc, by Thermo Fisher Scientific (3 mentions)
Cd11c pe cy7, by Thermo Fisher Scientific (2 mentions)
Cd11c fitc, by Thermo Fisher Scientific (2 mentions)
Cd8 percp efluor 710, by Thermo Fisher Scientific (2 mentions)
Cd3 pe cy7, by Thermo Fisher Scientific (2 mentions)
Mhc class 1 apc, by Thermo Fisher Scientific (2 mentions)
Granzyme b pe texas red, by BD (2 mentions)
Fc block, by Thermo Fisher Scientific (1 mentions)
Cd86 fitc, by BD (1 mentions)
Ccr7 pe, by BioLegend (1 mentions)
Cd40 pe, by BioLegend (1 mentions)
Cd11b apc, by BioLegend (1 mentions)
Cd11a pe, by BioLegend (1 mentions)
Cd18 fitc, by BD (1 mentions)
Lsrforetessa, by BD (1 mentions)
Saponin, by Merck Group (1 mentions)
Anisomycin, by Merck Group (1 mentions)
Z vad fmk, by Merck Group (1 mentions)
11 protocols using apc cd80
1
Multicolor Flow Cytometry Immunophenotyping
2
Monoclonal Antibodies for CHIKV Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mouse anti-CHIKV-nsP2 monoclonal antibody, used in this study was developed by us [58 (link)]. The anti-CHIKV-E2 monoclonal antibody was a kind gift from Dr. Manmohan Parida, DRDE, Gwalior, India. HRP linked secondary antibodies, H-2kd PE, I-A/I-E PE, isotype PE, isotype APC and HSP90 antibodies were purchased from BD Biosciences (San Jose, CA, USA). CD86 APC and CD80 APC were purchased from eBiosciences (San Diego, CA, USA). The monoclocal antibodies for cleaved caspase-3 (Asp175), cleaved caspase-8 (Asp387) and caspase-9 (C9) were purchased from cell signaling technology (Danvers, MA, USA). The anti-mouse Alexa Fluor 488 was purchased from Invitrogen (Carlsbad, CA, USA). Mouse IgG1 isotype control, GAPDH and β-actin were purchased from Abgenex India Pvt. Ltd. (Bhubaneswar, India). Saponin, Anisomycin and Bovine serum albumin fraction V were purchased from Sigma-Aldrich. 17-Allylaminogeldanamycin (17-AAG) and Z-VAD-FMK were purchased from Merck Millipore (Billerica, MA, USA).
3
Quantifying Tumor Immune Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect the in vivo immune response, 4T1 xenotransplant tumors were divided into eight groups and received treatments identical to in vivo therapeutic evaluation. On the 9th day, both primary tumors and distant metastases were harvested for single-cell suspensions fabrication. The prepared cells were further stained with CD11c-FITC (eBioscience, Catalog: N418), CD86-PE (eBioscience, Catalog: GL1), CD80-APC (eBioscience, Catalog: 16-10A1), F4/80-APC (Biolegend, Catalog: BM8), CD11b-PE (Biolegend, Catalog: M1/70), CD80-FITC (Biolegend, Catalog: 16-10A1), CD206-FITC (Biolegend, Catalog: C068C2), CD3-FITC (Biolegend, Catalog:17A2), CD8a-APC (Biolegend, Catalog: QA17A07) and CD4-PC5.5 (Biolegend, Catalog: RM4-5) antibody and then analyzed by flow cytometry. Serum was collected from different groups of mice, and cytokines including IL-6, IL-12, TNF-α and IL-10 were analyzed using ELISA kits according to the manufacturer's protocols. In addition, immunofluorescence staining was further conducted to investigate the infiltrating immune cells in tumor tissues.
4
Quantification of Antigen-Specific T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Analyses of labeled cell suspensions were performed on a custom Canto II (BD) and analyzed with FlowJo software (Tree Star, Inc.). Fluorochrome-conjugated mAbs used were CD11c—FITC (eBioscience, clone N418), CD86—PE (eBioscience, clone GL1), SIINFEKL-H-2Kb—PE (eBioscience, clone 25-D1.16) CD80—APC (eBioscience, clone 16-10A1), CD8α -APC (eBioscience, clone 53–6.7) and I-A/I-E—eFluor450 (MHC II; eBioscience, clone M5/114.15.2). The corresponding isotype controls were also purchased from eBioscience. A PE—dextran-conjugated multimer of SIINFEKL/H-2Kb was purchased from Immudex (Copenhagen, Denmark).
5
Immunotherapeutic Response Evaluation
6
Multiparameter Flow Cytometric Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The commercial sources for reagents were as follows: Antibodies used for flow cytometry were purchased from eBioscience (Live/Dead eFluor 506, CD45.2 Alexa Fluor 700, CD3 PE-Cy7, CD4 Pacific blue-eFluor 450, CD8 PerCP-efluor710, CD11b APC-eFluor 780, MHC Class I APC, CD40 APC, CD80 APC, CD86 APC), Invitrogen (granzyme B PE-Texas Red), and BD Pharmingen (CD11c-PE-Cy7). Murine anti-GM-CSF antibody was purchased from Thermo Fisher. DNAse I and Liberase TL were purchased from Roche. Recombinant murine GM-CSF protein was purchased from GenScript. Therapeutic anti-CTLA4 (clone 9H10 and 9D9) and anti-PD-L1 (clone 10F.9G2) were purchased from BioXcell.
7
Polymer-based Immunotherapeutic Delivery
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TLR7 agonist (R837), poly(D, l -lactic co-glycolic acid) (PLGA), polyvinyl alcohol (PVA), EDC/NHS and DAPI were provided by Sigma-Aldrich (St. Louis, USA). Hyaluronic acid (HA, MW = 50 kDa) were obtained from Bloomage Biotech Co., Ltd (Jinan, China). Type I collagen, hyaluronidase and collagenase D were supplied by Meilunbio Co., Ltd (Dalian, China). HSPC and CH were provided by AVT (Shanghai) Pharmaceutical Tech Co., Ltd. DOX (purity 99.0%) was supplied by Olympic Star Pharmaceutical Co., Ltd (Shenzhen, China). Poly(glycerin)10-monostearate (PG-C18) was supplied by Shandong Binzhou GIN&ING New Material Technology Co., Ltd. (Shandong, China). SA was provided by Changxing Pharmaceutical Co. Ltd (Huzhou, China). RPMI 1640 medium, FBS and penicillin-streptomycin solution were provided by Gibco (Carlsbad, USA). PE-CD11c, APC-CD40, APC-CD80, FITC-Gr-1, APC-CD86, FITC-CD3, PerCP-CD4 and APC-CD8 antibodies were obtained from eBioscience (San Diego, USA). ELISA kits of IL-6, IL-12, TNF-α, CXCL1, CCL2, IFN-γ, TGF-β and Arg-1 were supplied by R&D Systems (Minneapolis, USA). anti-CD62L, anti-MMP-9 and anti Arg-1 antibodies were supplied by BioXCell (West Lebanon, USA). Cell Counting Kit-8 (CCK-8) was obtained from Dojindo Laboratory (Kumamoto, Japan). All the other materials were immediately applied as supplied.
8
Multiparametric Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Surface and intracellular staining were performed as described previously using the indicated antibodies [33 (link)]. From BioLegend: Alexa fluor 488-Ly6C, PE-CD3, PE-CD86, APC-CD80, APC-I-A/I-F, Biotin-ICAM I, Biotin-H2kb, Alexa fluor 488-IFN-γ, Biotin-PD-L1, PE-Cy7-Ly6G, PE-CD34, PE-CD14, PE-F4/80, PE-Cy7-streptavidin, APC-streptatividin; From eBioscience: PE-CD4, FITC-IA/IE; From Raybiotech: FITC-class I H-2Kb; From BD bioscience pharmigen: v500-CD4, PE-CD8alpha, APC-CD11b, AF647-Ki67; from Southernbiotech: from Invitrogen: APC-CD11c; PE-streptavidin, FITC-streptavidin; from AbD Serotec: PE-CD62L; from R&D Systems: anti-human/mouse PE-Arginase 1. Viability was established by flow cytometry using the Fixable viability dye-eF780 from eBioscience. From Santa Cruz Biotechnology, NOS2-PE. When indicated, cells samples were treated overnight with 100 ng/ml lipololysaccharide (LPS) from Salmonella enterica serotype abortusequi (SIGMA).
9
Generation and Characterization of BMDCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMDCs were generated as noted (12 (link), 16 (link)). Briefly, BM cells were seeded in RPMI-1640 supplemented with 1% antibiotics/antimycotics and 10% heat-inactivated fetal calf serum (FCS) containing 20 ng/ml GM-CSF. A fresh complete medium change was performed on days 3 and 6. On day 8, nonadherent DCs were collected and resuspended in complete medium supplemented with PBS (to generate DCia) or 50 ng/ml IL-10 or 10 ng/ml LPS (to generate DC10, DClps) for 24 h. On day 9, cells were pulsed for 2 h at 37°C with 1 μM OVA (Sigma-Aldrich, Grade V; St. Louis, MO, USA) and then the cells were harvested (Supplementary Figure 1A
10
Splenocyte Isolation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Spleens were harvested aseptically and dissected from surviving mice. Cell suspensions were prepared by passage through nylon mesh, and red cells were removed with Lysing Buffer (BD). Splenocytes were resuspended as single cells in PBS and assessed with flow cytometry using the following antibodies: FITC-CD19, PE-CD86, APC-CD80, which were all purchased from eBioscience. The cells were stained with these antibodies for 30 min at 4°C in the dark. Isotype control antibodies (eBioscience) were used to control for non-specific antibody binding to the cells. Cells were then washed, resuspended in PBS, and analysed with a BD FACSCanto II flow cytometer. Data were analysed with FlowJo7.6 software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!