Lymphocytes were isolated as described and stained with anti-mouse IgD PerCP-Cy5.5 (1:400, 405710), LPAM (integrin α4β7) PE (1:100, 120605; (Biolegend, San Diego, CA), B220 V500 (1:400, 561227), CD19 APC-H7 (1:200, 560143), CD80 PE (1:500, 553769), CD273 APC (1:200, 560086), CD138 PE (1:200, 553714), IgM PE-Cy7 (1:200, 552867; BD Biosciences), CCR9 PE-Cy7 (1:100, 25-1991), CD73 PE-Cy7 (1:50, 25-0731), IgA PE (1:50, 12-4204), GL7 eFluor 450 (1:100, 48-5902), CD38 Alexa700 (1:800, 56-0381), CD21/35 Pacific Blue (1:800, 57-0212; eBiosciences) or CCR10 APC (1:100, FAB2815A; R and D systems. Minneapolis, MN) and were analysed using LSR II (BD Biosciences) or Navios (Beckman Coulter, Brea, CA) flow cytometers. For sorting, cells were labelled with anti-mouse CD138 PE (1:200, 553714), CD19 PE-Cy7 (1:200, 552854), CD80 APC (1:200, 560016) and GL7 FITC (1:100, 553666) before sorting using a FACSAria (BD Biosciences). Cells were sorted into tubes that had been coated with 2% BSA/PBS overnight, and pelleted by centrifugation at 600 g before being resuspended in PBS and injected into recipient mice or used for gene sequence analysis.
Cd80 apc
Manufactured by BD
Sourced in United States
The CD80-APC is a fluorescent-labeled antibody that binds to the CD80 cell surface protein. It is commonly used in flow cytometry applications to identify and analyze cells expressing CD80.
Automatically generated - may contain errors
Lab products found in correlation
Cd80 pe, by BD (2 mentions)
Cd64 apc, by BD (2 mentions)
Cd86 pe, by BD (2 mentions)
Cd11c fitc, by BD (2 mentions)
Cd4 apc, by BD (2 mentions)
Cd11b fitc, by BD (2 mentions)
F4 80 pe, by BD (2 mentions)
Ifn γ pe, by BD (2 mentions)
Cd86 pe cy7, by BD (2 mentions)
High molecular weight poly vinyl alcohol pva, by Thermo Fisher Scientific (2 mentions)
Cd40 pe, by BD (2 mentions)
Cd138 pe, by BD (1 mentions)
Gl7 fitc, by BD (1 mentions)
Igd percp cy5, by BioLegend (1 mentions)
Ccr10 apc, by R&D Systems (1 mentions)
Cd19 apc h7, by BD (1 mentions)
Cd273 apc, by BD (1 mentions)
Igm pe cy7, by BD (1 mentions)
Cd73 pe cy7, by Thermo Fisher Scientific (1 mentions)
Iga pe, by Thermo Fisher Scientific (1 mentions)
9 protocols using cd80 apc
1
Lymphocyte Isolation and Characterization
2
Multiparametric FACS Analysis of RTMs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RTMs were collected at 5 × 105 per FACS tube, washed with FACS buffer (1% BSA/5 mM EDTA/PBS, pH 7.2), and incubated in blocking buffer (1:100 anti-CD16/CD32 in FACS buffer) for 15 min at 4°C. After blocking, RTMs were washed with FACS buffer and stained by anti-CD45-BV711 (BD), F4/80-PE (BioLegend), CD11b-FITC (BD), Siglec-F-PE (BD), CD11c-APC-Cy7 (BioLegend), or CD80-APC (BD) at a dilution of 1:200 by FACS buffer for 30 min at 4°C. After washing with FACS buffer, cells were analysed by LSRFortessa X-20 (BD) and FlowJo software, v 10.8.1.
3
Multiparameter Flow Cytometry Immunophenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were digested into single cells and suspended in PBS with 2% FBS. The cell suspension was incubated with the antibody for 30 min at 4°C. After the incubation, the expression of the markers in this study were analyzed by the CytoFLEX Flow Cytometer (Beckman); for cell sorting, cells were sorted by the Moflo (Beckman), and then analyzed using FlowJo (Treestar). The following antibodies were purchased from BD Biosciences: CD11b-APC-Cy7, CD34-PE, CD43-APC, CD14-FITC, CD80-APC, CD86-APC, CD163-PE, CD206-PE, CD64-APC, HLA-DR-APC, CD25-PE, CD69-APC. LAG3-PE, CD3-V500, CD4-FITC, and CD8-PE.
4
Multiparametric Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood, spleens, and mesenteric lymph nodes (mLN) were collected 24 h after CLP surgery or sham surgery to obtain single cells for flow cytometry. Cells were stained for 30 min at 4°C with the following antibodies: CD45-Percp, CD3-FITC, CD4-Percp, CD8-APC, B220-PE, CD11c-APC, CD11b-FITC, F4/80-PE, and Ly6G-PE. Following incubation, red blood cells were lysed and washed 3 times with a FACS buffer before collecting data. Immunophenotyping analysis of the BMDMs was conducted using a flow cytometry technique. Briefly, 100 μl of cell suspension was incubated for 5 min with 5 μl of Fc blocker (Innovex, USA). Then, 1 μl of monoclonal antibodies (CD11b-FITC, F4/80-PE, F4/80-FITC, MHCII-FITC, CD80-PE, CD80-APC, CD16/32-Percp-cy5.5, and CD64-APC, BD, USA) were added and incubated at 4°C for 30 min and washed 3 times with the FACS buffer. All samples were analyzed using a FACSCalibur flow cytometer (BD, USA).
5
Influenza Vaccine Immunogenicity Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AEAR was prepared in our lab [22 (link)]. 2014/2015 seasonal influenza virus split vaccine was purchased commercially from Shenzhen sanofi Pasteur biological products co., LTD (Shenzhen China). Goat anti-mouse peroxidase conjugates (IgG-HRP, IgG1-HRP and IgG2a-HRP) were purchased from Southern Biotech Inc. USA. Concanavalin A (ConA), lipopolysaccharide (LPS, from Escherichia coli 055:B5) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were from Sigma-Aldrich Co. LLC. USA. Imject Alum Adjuvant (Alum) was from Thermo Scientific Pierce USA. CD11c-FITC, CD3-PE, CD4-APC, CD8a-FITC, CD44-PE, CD25-APC, Foxp3-PE, IFN-γ-PE, IL-4-PE, CD86-PE, CD40-APC, CD80-APC, MHC-II-PE, Cytofix/Cytoperm and Perm/Wash buffer were purchased from BD Bioscience USA. Carboxyfluorescein diacetate succinimidyl ester (CFSE), Treg staining kit were from eBioscience USA. All the other reagents were analytically purified in China.
6
Nanoparticle-based Drug Delivery Platform
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Soy Lecithin and cholesterol were obtained from Avanti. PTX was obtained from Aladdin Chemical Co., Ltd. Sulfhydryl polyethylene glycol (SH‐PEG2000) and DSPE‐PEG‐FITC were purchased from Shanghai ToYong Bio Tech. Inc. Chloroauric acid trihydrate (HAuCl4·3H2O) was purchased from Dibo Biological Technology Co., Ltd. Hexadecyl trimethyl ammonium bromide (CTAB) Shanghai Mokai Biological Technology Co., Ltd. Enhanced BCA Protein Assay Kit, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT), and penicillin–streptomycin solution were purchased from Beyotime. 4′,6‐diamidino‐2‐phenylindole (DAPI), Dulbecco's modified Eagle medium (DMEM) medium with high glucose, and RPMI1640 medium were purchased from HyClone. 3,3′‐dioctadecyloxacarbocyanine perchlorate (DiO) was obtained from Protein Biological Technology Co., Ltd. (Nanjing, China). Alexa Fluor 647 anti‐CD9 antibody was purchased from Abcam. CD11c‐FITC, CD3‐FITC, CD4‐APC, CD8‐Percp‐Cy5.5, CD80‐APC, and CD86‐PE antibodies were purchased from BD Pharmingen. ELISA kits for interlekin‐6 (IL‐6), interferon‐γ (IFN‐γ), and tumor necrosis factor‐α (TNF‐α) were obtained from Multi Sciences Biotech, Co., Ltd. The other chemical reagents with analytical grade were obtained from Solarbio Life Sciences.
7
Immunomodulatory Nanoparticles for T-cell Therapy
8
Antigen-specific Immunomodulation Using PLGA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MOG peptide (MOG35-55, MEVGWYRSPFSRVVHLYRNGK) was synthesized by Genscript (Piscataway, NJ, United States) with >98% purity. Rapamycin was purchased from LC Laboratories (Woburn, MA, United States). 50:50 poly(lactic-co-glycolide) (PLGA) was purchased from LACTEL Absorbable Polymers. High molecular weight poly(vinyl alcohol) (PVA) was purchased from Alpha Aesar (Tewksbury, MA, United States). Dichloromethane (DCM) was purchased from Sigma Aldrich (St. Louis, MO, United States). Dimethyl sulfoxide (DMSO), micro-bicinchoninic acid (mBCA) assay, and eBioscience FoxP3 Fixation/Permeation kit were purchased from Thermo Fisher Scientific (Waltham, MA, United States). Antibodies for flow cytometry, including BV605-CD11c, v450-CD25, PE-T-bet, PE-CD40, PE-Cy-7-CD4, PE-Cy-7-CD86, APC-FoxP3, APC-CD80 were purchased from BD Biosciences. Zombie NIR Fixable Viability Kit was purchased from BioLegend.
9
Tumor Macrophage Isolation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse tumor tissues were added with 5–10 mL HBSS + Ca/Mg (PB180323, Procell) + 10 μg/mL DNase I (10104159001, Sigma-Aldrich) + 25 μg/mL Liberase (5401054001, Roche), detached at 37°C for 30 min and vortexed once every 10 min. After lysis, the tumor suspension was filtered with a 70 m filter, placed on ice, washed with pre-cooled PBS, and centrifuged at 4°C and 1000 × g for 5 min. Blood cells were lysed in ACK lysis buffer (A1049201, Invitrogen) for 10 min, which was halted by PBS, followed by centrifugation at 4°C and 1000 × g for 5 min. The samples were resuspended in FACS buffer, blocked with rat serum IgG (1 μg/106 cells, I4131-10 MG, Sigma-Aldrich) for 15 min and incubated with the following antibodies (all from BD Bioscience): BV480-TCRβ (746385), FITC-CD11b (561688), PE-F4/80 (565410), Alexa Fluor® 647-CD206 (565250) and APC-CD80 (560016). For all channels, the positive and negative cells were gated on the basis of the FMOs.
Macrophage panel:
M1 macrophages: Hoechst-, TCRβ-, CD11b+, F4/80+, CD80+ and CD206-.
M2 macrophages: Hoechst-, TCRβ-, CD11b+, F4/80+, CD80-, and CD206 + .
Macrophage panel:
M1 macrophages: Hoechst-, TCRβ-, CD11b+, F4/80+, CD80+ and CD206-.
M2 macrophages: Hoechst-, TCRβ-, CD11b+, F4/80+, CD80-, and CD206 + .
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!