Primary neuronal and microglial cultures grown in 15 mm glass coverslips were fixed by incubation with 4% formaldehyde (15 minutes), followed by incubation with 1:100 Bgtx-Alexa-555 (to label α7, Invitrogen, Eugene, OR) and 1:500 CD90-FITC (to label neurons, Becton Dickinson, CA) or CD80-PE (to label microglia, Becton Dickinson, CA) for 1hr at room temperature in dark. Cells were then washed 3 times with PBS 1X(pH 7.2), coverslips placed with mounting media containing DAPI (to label cellular nuclei), and fluorescence acquired by confocal imaging (Zeiss LSM Meta 510; Carl Zeiss, Pleasanton, CA) at the Confocal Imaging Facility, University of Puerto Rico (http://cifupr.org/ ) under a 40X magnification.
Cd80 pe
Manufactured by BD
Sourced in United States
CD80-PE is a fluorescently labeled antibody used to detect the CD80 cell surface protein. CD80 is a co-stimulatory molecule expressed on antigen-presenting cells, which plays a role in T cell activation. The PE (phycoerythrin) fluorescent tag allows for the identification and analysis of CD80-positive cells using flow cytometry techniques.
Automatically generated - may contain errors
Lab products found in correlation
Cd86 pe, by BD (15 mentions)
Cd11c apc, by BD (13 mentions)
Cd86 fitc, by BD (12 mentions)
Cd86 apc, by BD (10 mentions)
Facscanto 2, by BD (7 mentions)
Cd83 apc, by BD (6 mentions)
Hla dr fitc, by BD (6 mentions)
Facscalibur flow cytometer, by BD (6 mentions)
Hla dr percp, by BD (5 mentions)
Cd14 fitc, by BD (5 mentions)
Cd3 fitc, by BD (5 mentions)
Ccr7 pe, by BD (5 mentions)
Cd40 pe, by BD (5 mentions)
Cd4 fitc, by BD (4 mentions)
Cd8 fitc, by BD (4 mentions)
Lipopolysaccharides (lps), by Merck Group (4 mentions)
7 aad, by BD (3 mentions)
Cd56 pe, by BD (3 mentions)
Cd4 percp, by BD (3 mentions)
Cd14 apc, by BD (3 mentions)
60 protocols using cd80 pe
1
Immunofluorescence Labeling of Neurons and Microglia
2
Phenotyping of Cultured Primary Immature Dendritic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The phenotype of the cultured primary imDCs was confirmed at day 6 using flow cytometry analysis, essentially as described before [27] (link), [29] (link). The cells were analyzed using the primary labeled antibodies Lineage1-FITC, CD11c-APC, HLR-DR-V500, CD80-PE, CD83-PECy7 and CD86-V50 (Becton Dickinson). DC-SIGN levels of imDCs, Raji wild type, and Raji DC-SIGN cells were determined using an anti-DC-SIGN antibody and a secondary PE-labeled antibody (both R&D systems, MN, USA). Flow cytometry analysis was performed on a LSR-II (Becton Dickinson). Data was analyzed using Kaluza 1.2.
3
Quantification of DC Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used the following monoclonal antibodies reactive against DC markers: CD86-PE, HLA-DR-PerCP, CD80-PE, CCR2-AlexaFlour 647, CCR5-PerCP (Becton Dickinson, Heidelberg, Germany) and respective monoclonal antibody isotype controls. Cells were collected, washed once with PBS, blocked with PBS containing mouse IgG (50 μg ml−1) incubated on ice for 15 min and stained with the corresponding monoclonal antibodies for 30 min. Cells were washed and resuspended in cell-fix solution (Becton Dickinson). Cells were then analyzed using a FACSCalibur cytometer (Becton Dickinson). Acquisition was performed using CellQuest software and analyses were performed using Flowjo software (Treestar Inc., Ashland, OR, USA).
4
Immunotherapeutic Agents and Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
WT-1, BIRC5, MUC-1, and TERT were purchased from MyBioSource (San Diego, CA, USA). Recombinant mGM-CSF was purchased from PeproTech (Cranbury, NJ, USA). Cisplatin and lipopolysaccharide (LPS) were purchased from Sigma-Aldrich (Burlington, MA, USA). Fluorophore-labeled monoclonal antibodies (Annexin V-FITC, 7-AAD, CD3e-FITC, CD4-PE, CD8-Pacific blue, CD11c-APC, CD80-PE, CD86-FITC, Fixable Viability Stain 780, IFN-γ-BV605, MHC I-PE, MHC II-PerCP-Cy5.5) were purchased from BD Bioscience (Franklin Lakes, NJ, USA). Cell trace violet was purchased from Invitrogen (Waltham, MA, USA). LLC1 was purchased from ATCC (Manassas, VA, USA).
5
Phenotyping of Immune Cells by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies for surface markers staining of DCs and T cells were obtained from BD Biosciences, New York, USA (anti-human CD3-PE, CD3-FITC, CD8-PerCP, CD8-APC, CD56-PE, NKG2D-APC, CD4-FITC, CD4-PerCP, CD107a-FITC, CD25-APC, CD45RO-FITC, CD27-PerCPCY5.5, CD57-APC, CCR7-PE, CD14-APC, CD80-PE, CD83-APC, CD86-FITC, HLA-DR-FITC). Antibodies for intracellular proteins staining were also obtained from BD Biosciences (anti-human IFNγ-APC, TNFα-PECY7, granzyme B-FITC, FoxP3-PE). The intracellular staining was performed by fixing and permeabilizing cells with Cytofix/Cytoperm (BD Biosciences). The experiments were performed by using FACS CantoII (BD Biosciences) flow cytometer, and data were analyzed by using the Flowjo software.
6
Zika Virus Detection in Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed with paraformaldehyde (PFA) and treated with either BSA (extracellular staining) or BSA/Saponin to measure intracellular staining. The following antibodies were used to detect Zika virus: anti-Flavivirus 4g2 mouse IgG2a (NovusBio), anti-Flavivirus 4g2 monoclonal rabbit (Absolute Antibody), anti-Zika virus monoclonal Rabbit (Genetex).
All other antibodies were anti-human: DC-SIGN mouse IgG1 (AZN-D1), anti-langerin mouse IgG1 (10E2) both in house made, PE conjugated CD207 (langerin), APC conjugated CD1a (BD Biosciences), CD86-FITC (BD Pharmingen), CD80-PE (BD Pharmingen), CD83-APC (BD Pharmingen), DC-SIGN-FITC (R&D systems), CD3-APC/Fire750 (Biolegend), CD11c-APC (Biolegend), PEcy7-HLA-DR (BD Pharmingen), APCcy7-CD14 (BD Biosciences), APCcy7-CD11c (Biolegend),APC-AXL (Thermofisher, PE-MerTK (Thermofisher) and anti-Tyro3 (Thermofisher). For secondary detection the following antibodies were used: AF488-conjugated goat anti-mouse IgG2a (Invitrogen), AF647-conjugated donkey anti-rabbit (Thermofisher), FITC-conjugated goat-anti-mouse IgM (Invitrogen), AF488-conjugated donkey anti-rabbit (Thermofisher).
Flow cytometric analyses were performed on a BD FACS Canto II (BD Biosciences) and data was analysed using FlowJo V10 software (TreeStar).
All other antibodies were anti-human: DC-SIGN mouse IgG1 (AZN-D1), anti-langerin mouse IgG1 (10E2) both in house made, PE conjugated CD207 (langerin), APC conjugated CD1a (BD Biosciences), CD86-FITC (BD Pharmingen), CD80-PE (BD Pharmingen), CD83-APC (BD Pharmingen), DC-SIGN-FITC (R&D systems), CD3-APC/Fire750 (Biolegend), CD11c-APC (Biolegend), PEcy7-HLA-DR (BD Pharmingen), APCcy7-CD14 (BD Biosciences), APCcy7-CD11c (Biolegend),APC-AXL (Thermofisher, PE-MerTK (Thermofisher) and anti-Tyro3 (Thermofisher). For secondary detection the following antibodies were used: AF488-conjugated goat anti-mouse IgG2a (Invitrogen), AF647-conjugated donkey anti-rabbit (Thermofisher), FITC-conjugated goat-anti-mouse IgM (Invitrogen), AF488-conjugated donkey anti-rabbit (Thermofisher).
Flow cytometric analyses were performed on a BD FACS Canto II (BD Biosciences) and data was analysed using FlowJo V10 software (TreeStar).
7
Tumor-Infiltrating Immune Cell Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
M-chlorin e6 PDT was performed as described above. Mice were euthanized by cervical dislocation on day 2 after LED irradiation. The tumors were removed and treated with 10% collagenase (Wako), shredded using a gentleMACS™ Dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany), and passed through a 70-μm filter. Next, 1.0 × 106 cells were mixed with 1 μL of Fc-block/100 μL of stain buffer (BD Biosciences, San Jose, CA) and left on ice for 10 min. Subsequently, 1 μL of fluorescent antibody (CD11b-APC [BD Biosciences], CD206-BV421 [Biolegend, San Diego, CA], CD80-PE [BD Biosciences], CD86-PE-Cy7 [BD Biosciences], CD45-PE-Vio®770 [Miltenyi Biotec], CD3-FITC [Miltenyi Biotec], CD8a-APC-H7 [BD PharMingen, San Diego, CA], CD127-APC [BD PharMingen], CD25-PE [BD Biosciences], or CD4-BV421 [Biolegend]) diluted with 100 μL of stain buffer was added and left on ice for 30 min. The cells were then washed with 500 μL of phosphate-buffered saline (PBS) and suspended in 500 μL of stain buffer. Next, 5 μL of 7-AAD (BD Biosciences) was added to prepare a sampling solution, which was analyzed on a BD FACSVerse™ (BD Biosciences).
8
Comprehensive Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The monoclonal antibodies used in this study included CD3 PerCP, CD5 APC-R700, CD8-FITC, CD11c PE-Cy5, CD14 APC-H7, CD19 PE-Cy5, CD25 PE-Cy7, CD24 FITC, CD27 PE-Cy7, CD28 PE-Cy5, CD38 APC-H7, CD45 V500, CD45RO APC, CD56 APC, CD57 FITC, CD80 PE, CD86 APC, CD123 APC, CD127 BV421, CCR7 PE, TIGIT BV421, HLA DR V450, TIM3 PE, CXCR5 APC-R700, PD1 BV421, and PDL1 PE-Cy7 (BD Biosciences).
9
Characterization of Bone Marrow-Derived Dendritic Cells
10
Phenotypic Analysis of Langerhans Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immature and mature LCs were phenotyped using CD1a-APC (BD Pharmingen), langerin-PE (Novocastra), CD86-FITC (BD Pharmingen), CD80-PE (BD Pharmingen), CD83-APC (BD Pharmingen), CD4-AF488 (Biolegend), CD195 (CCR5)-PE (BD Pharmingen) and unlabeled CXCR4 (R&D systems) for which secondary detection with Goat-anti-Mouse-A488 (Invitrogen) was used. HIV-1 infection and transmission samples were stained for CD1a-APC (LC marker), CD3-PerCP (T cell marker), and p24-PE (HIV-1 envelope protein, Beckman Coulter). Immature vaginal LCs were further sorted with a FacsARIA 3 laser sorter (BD Biosciences) after staining for CD1a-APC into CD1a positive and CD1a negative fractions. Samples were analysed using FACSCanto II flow cytometers (BD Biosciences) and data analysis was carried out with FlowJo V10.
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!