The following antibodies were purchased from BioLegend, eBioscience, or BD Biosciences, and used for flow cytometry or enrichment of specific cell types: FITC-Sirpα, PE/Cy7-Sirpα PerCP/Cy5.5-B220, PE/Cy7-CD11c, Brilliant Violet 605-CD11c, Pacific Blue-IA/IE, FITC-YAe, PerCP/Cy5.5-CD45.1, APC-CD45.2, PE/Cy7-CD86, APC-CD86, PE-PD-L1, APC-CD200, PE/Cy7-CD40, PE-Foxp3, Pacific Blue-Foxp3, PerCP/Cy5.5-Thy1.1, APC-TCRβ, FITC-CD8α, Alexa700-CD8α, Alexa700-CD4, Brilliant Violet 605-CD4, Brilliant Violet 605-CD25, APC-CD25, PE/Cy7-CD3, PE/Cy7-CD24, Propidium iodide solution, Fixable Viability Dye eFluor780, Biotin-CD8β, Biotin-CD4, Biotin-CD25, and Biotin-B220. AlexaFluor647-conjugated OVA and crimson bead (0.02 μm) were purchased from ThermoFisher Scientific. Streptavidin-RapidSpheres isolation kit and CD11c-MicroBeads were purchased from STEMCELL technologies and Miltenyi Biotec, respectively.
Cd40 pe cy7
Manufactured by BD
Sourced in United States
The CD40-PE-Cy7 is a fluorescently labeled monoclonal antibody that binds to the CD40 surface antigen. It is a tool for the identification and analysis of CD40-expressing cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Cd11c pe cy7, by BD (2 mentions)
Cd68 pe, by BD (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Propidium iodide solution, by BD (1 mentions)
Cd86 apc, by BD (1 mentions)
Apc tcr β, by BD (1 mentions)
Cd86 pe cy7, by BD (1 mentions)
Alexa fluor 647 conjugated ova, by Thermo Fisher Scientific (1 mentions)
Cd25 apc, by BD (1 mentions)
Fixable viability dye efluor 780, by BD (1 mentions)
Biotin cd4, by BD (1 mentions)
Foxp3 pe, by BD (1 mentions)
Fitc sirpα, by BD (1 mentions)
Brilliant violet 605 cd11c, by BD (1 mentions)
Pacific blue ia ie, by BD (1 mentions)
Cd45 apc, by BD (1 mentions)
Pacific blue foxp3, by BD (1 mentions)
Percp cy5 5 thy1, by BD (1 mentions)
Cd8α fitc, by BD (1 mentions)
Alexa700 cd8α, by BD (1 mentions)
7 protocols using cd40 pe cy7
1
Multiparameter Immune Cell Analysis
2
Multiparameter Flow Cytometry Panel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A single-cell suspension was blocked with mouse FcR blocking reagent (Miltenyi Biotec, USA) at 4°C for 10 min prior to surface staining. For intracellular IFN-γ staining, the cells were fixed and permeabilized with Fixation/Permeabilization Kit (BD Biosciences, USA) and then stained with IFN-γ antibody. The following anti-mouse antibodies were used: PE-CD44 (clone:IM7), APC-CD24 (clone:M1-69), FITC-CD3e (clone: 145-2C11), PE-CD45 (clone:30-F11), APC-MHC class II (I-A/I-E) (clone: AMS-32.1), PE-CD86 (clone: GL 1) from eBioscience (San Diego, CA, USA); PE/Cy7-CD8a (clone: 53-6.7), PE-MHC class I (H-2Kd) (clone: AMS-32.1), PE/Cy7-CD40 (clone: 3-23), PE/Cy7-CD11c (clone: HL3) from BD Biosciences (USA); and APC-CD8a (clone: 53-6.7), PE-IFN-γ (clone: XMG 1.2), APC-CD11c (clone:N418), APC-CD80 (clone: 16-10A1) from Biolegend (USA). For ALDH1 staining, the ALDH1 activity was detected by the ALDEFLUOR kit (Stem Cell Technologies, Canada) according to the instructions of the manufacturer. Data were acquired on BD FACS Calibur (USA) and analyzed by the Flow-Jo software (Tree Star, OR) version 7.1.2. Fixable viability dye eFluor 520 (eBioscience, USA) was used to exclude dead cells.
3
Cryopreserved PBMC Characterization and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cryopreserved PBMC were thawed and allowed to rest overnight (37°C, 5% CO2) as previously described [41 (link), 42 (link)]. Plating of cells (1x106), staining for viability, bacteria binding (50:1—bacteria:cells ratio [43 (link)]), blocking (human IgG -25 μl of a 1 mg/ml solution; mouse IgG -25 μl of a 200 μg/ml solution) and staining of surface targets with monoclonal antibodies were performed as described in detail in [40 (link)]. Monoclonal antibodies (mAbs) against the following molecules were used: CD19-ECD (clone J3-119; Beckman Coulter -BC-), CD38-PE-Cy5 (clone LS1298-4-3; BC), CD14-QDot 655 (clone TuK; Invitrogen), CD21-BV711 (clone B-ly4; Becton-Dickinson -BD-), integrin α4β7-Alexa647 (clone ACT-1; Millennium, The Takeda Oncology Co), CD3-Alexa Fluor 700 (clone UCHT1; BD), IgD-FITC (polyclonal goat anti-sera; Southern Biotech), CD27-PE (clone L128; BD), CD40-PE-Cy7 (clone 5C3; BD), IgA-Biotin (polyclonal goat anti-sera; Southern Biotech) and Pacific Orange-Streptavidin (Invitrogen, USA). Finally, stained cells were fixed with 1% PFA in PBS until data collection in a LSRII (BD) instrument.
4
Investigating Immune Cell Responses to TLR Agonists
5
Flow Cytometry Characterization of WJ-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Control and ischemic WJ-MSCs were characterized for surface antigen expression by flow cytometry analysis. Anti-human antibodies such as CD90-PE, CD73-PE, CD105-PE, CD34-PE, CD80-PE, and CD40-PECy7 were used to mark the cell surface markers (all from BD Pharmingen, SanDiego, CA, USA). Mouse isotype antibodies served as control (BD Pharmingen). At least 10,000 events were acquired on BD FACSCalibur flow cytometer and analyzed with BD CellQuest Pro software.
6
Quantifying Macrophage Subpopulations in Bone Marrow
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow samples were obtained by standard bone marrow puncture using sterile heparin anticoagulant tubes. Bone marrow samples were filtered using flow cytometry tubes. CD14-FITC (Cat No.: 555397), CD68-PE (Cat No.: 565595), CD64-APC (Cat No.: 561189), CD40-PEcy7 (Cat No.: 561215), CD206-PE (Cat No.: 555954), CD163-PEcy7 (Cat No.: 556018), and isotype control antibodies (BD Biosciences, USA) were added to the tubes. The samples were then stained for 15 min in the dark at room temperature. After red blood cell lysis, the cells were washed with PBS. Finally, the cells were detected using a FACSCalibur flow cytometer (BD Biosciences, USA). Data analysis was performed using the Cell Quest software (Becton Dickinson, version 3.1).
Macrophages were defined as CD14+CD68+ cells. M1 macrophages were defined as CD64+CD40+ macrophages. M2 macrophages were defined as CD206+CD163+ macrophages (detail in Supplemental Figure1 ).
Macrophages were defined as CD14+CD68+ cells. M1 macrophages were defined as CD64+CD40+ macrophages. M2 macrophages were defined as CD206+CD163+ macrophages (detail in Supplemental Figure
7
Flow Cytometry Analysis of DC Phenotypes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human DC phenotypes were assessed via flow cytometry (Becton Dickinson, LSRFortessa). Cells were incubated with fluorochrome-conjugated monoclonal antibodies (CD40-PE-Cy7, CD86-BV510, HLA-DR-APC, CD83-APC-Cy7, CD274-APC, and CD54-PE; BD PharMingen) for 30 min on ice, washed, and analyzed using FlowJo software as previously described [44 ].
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!