The flow cytometry was carried out using the BD FACSCanto™ II Cell Analyzer (BD Bioscience). The analysis of THP-1 macrophages, human classical monocytes, and Kupffer cells was performed as described previously with some modifications (Tian et al., 2016 (link)). Briefly, cells were collected and incubated with primary antibodies IgG2a-PE, IgG1-APC, CD14-PE, APC anti-human CD11b antibody (Biolegend, #301310), APC anti-human CD16 antibody (Biolegend, #360705), F4/80-PE (eBioscience, #12-4801-82), or IgG2a-PE (eBioscience, #12-4321-80) at 4°C for 30 minutes. Cells were rinsed with PBS three times before the flow cytometry analysis. Data were analyzed by FlowJo 10 software (BD Bioscience)
Apc anti human cd16 antibody
Manufactured by BioLegend
The APC anti-human CD16 antibody is a fluorochrome-conjugated monoclonal antibody that binds to the CD16 antigen expressed on human cells. CD16, also known as FcγRIII, is a low-affinity Fc receptor that plays a role in antibody-dependent cell-mediated cytotoxicity (ADCC). This antibody can be used in flow cytometry applications to identify and characterize CD16-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2 cell analyzer, by BD (2 mentions)
F4 80 pe, by Thermo Fisher Scientific (2 mentions)
Cd14 pe, by BioLegend (2 mentions)
Igg2a pe, by BioLegend (2 mentions)
Anti human cd11b apc antibody, by BioLegend (2 mentions)
Igg2a pe, by Thermo Fisher Scientific (2 mentions)
Igg1 apc, by BioLegend (2 mentions)
Sepmate 15 tube, by STEMCELL (1 mentions)
Histopaque 1077, by Merck Group (1 mentions)
K2edta, by BD (1 mentions)
Apc cyanine7 anti human cd3 antibody, by BioLegend (1 mentions)
Fitc anti human cd4 antibody, by BioLegend (1 mentions)
Streptavidin bv421, by BioLegend (1 mentions)
Human fc block, by BD (1 mentions)
Viability dye, by Thermo Fisher Scientific (1 mentions)
Pe cy7 anti human cd4 antibody, by BioLegend (1 mentions)
Fitc anti human cd19 antibody, by BioLegend (1 mentions)
Apc cy7 anti human cd3 antibody, by BioLegend (1 mentions)
Fitc anti human cd69 antibody, by BioLegend (1 mentions)
4 protocols using apc anti human cd16 antibody
1
Flow Cytometry Analysis of Macrophages
2
Flow Cytometry Analysis of Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The flow cytometry was carried out using the BD FACSCanto™ II Cell Analyzer (BD Bioscience). The analysis of THP-1 macrophages, human classical monocytes, and Kupffer cells was performed as described previously with some modifications (Tian et al., 2016 (link)). Briefly, cells were collected and incubated with primary antibodies IgG2a-PE, IgG1-APC, CD14-PE, APC anti-human CD11b antibody (Biolegend, #301310), APC anti-human CD16 antibody (Biolegend, #360705), F4/80-PE (eBioscience, #12-4801-82), or IgG2a-PE (eBioscience, #12-4321-80) at 4°C for 30 minutes. Cells were rinsed with PBS three times before the flow cytometry analysis. Data were analyzed by FlowJo 10 software (BD Bioscience)
3
Isolation and Characterization of Immune Cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cryopreserved CD3+ Pan T cells (PB009-1F-C-5M), CD19+ B cells (PB010-P-F-C), CD14+ monocytes (PB-011-P-F-1-C), and CD56+ natural killer cells (PB012-P-F-C) were purchased from AllCells (Shanghai, China). The CD3+ Pan T cells were stained with APC/Cyanine7 anti-human CD3 Antibody (300425, BioLegend), FITC anti-human CD4 Antibody (300506, BioLegend) and PE anti-human CD8 Antibody (344705, BioLegend) following the manufacturer’s instructions. Then, the Pan T cells were sorted into CD4+ T cells and CD8+ T cells with a BD FACSAria III instrument. CD14+ monocytes were stained with PE anti-human CD14 Antibody (301805, BioLegend) and APC anti-human CD16 Antibody (302011, BioLegend) following the manufacturer’s instructions. CD14+ monocytes were further separated into CD16+ monocytes and CD14+ monocytes (CD16-) with a BD FACSAria III instrument.
For fresh PBMCs, venous blood was collected from a healthy donor into a plastic blood tube spray-coated with K2EDTA (367863, Becton Dickinson, NJ, USA). Blood was dissolved in an equal volume of 1 x DPBS solution and added to a SepMate™-15 tube (86415, StemCell Technologies, Canada) containing Histopaque-1077 (10771-100 ml, Sigma-Aldrich, MO, USA). After centrifugation (1200 x g, 10 min), PBMCs were collected, washed twice, and resuspended with a 1 x DPBS solution.
For fresh PBMCs, venous blood was collected from a healthy donor into a plastic blood tube spray-coated with K2EDTA (367863, Becton Dickinson, NJ, USA). Blood was dissolved in an equal volume of 1 x DPBS solution and added to a SepMate™-15 tube (86415, StemCell Technologies, Canada) containing Histopaque-1077 (10771-100 ml, Sigma-Aldrich, MO, USA). After centrifugation (1200 x g, 10 min), PBMCs were collected, washed twice, and resuspended with a 1 x DPBS solution.
4
Comprehensive Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flow cytometry was performed with MACS Quant with the following antibodies: human BD Fc Block (BD Biosciences, Franklin Lakes, New Jersey), Viability Dye (eBioscience, San Diego, CA), Pacific Blue anti-human CD14 Antibody (BioLegend, San Diego, CA), FITC anti-human CD19 Antibody (BioLegend, San Diego, CA), PE-Cy7 anti-human CD56 Antibody (BioLegend, San Diego, CA), APC-Cy7 anti-human CD3 Antibody (BioLegend, San Diego, CA), APC anti-human CD16 Antibody (BioLegend, San Diego, CA), BV421 anti-human CD25 Antibody (BioLegend, San Diego, CA), FITC anti-human CD69 Antibody (BioLegend, San Diego, CA), PerCP anti-human CD8a Antibody (BioLegend, San Diego, CA), PE-Cy7 anti-human CD4 Antibody (BioLegend, San Diego, CA), Biotin Rabbit Anti-Mouse FMC63 scFv (BioSwan, Shanghai, China), and BV421-Streptavidin (BioLegend, San Diego, CA).
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!