Exosomes were characterized based on the expression of exosomes markers by flow cytometry. Extracted exosomes (10 μg) were incubated with 1 µl Aldehyde/Sulfate Latex Beads (Invitrogen) 15 min at RT. After washed, exosomes-beads complex were incubated with Anti-CD63-PE, Anti-CD81-APC, Anti-CD9-PE (Biolegend). The morphology of exosomes was observed using the transmission electron microscopy (TEM). About 20 μl exosome suspensions were dropped on a sample-loaded copper mesh. Samples were negatively stained with 2% uranyl acetate solution and air dried for 30 min.
Anti cd81 apc
Manufactured by BioLegend
Anti-CD81-APC is a monoclonal antibody that specifically binds to the CD81 antigen. CD81 is a cell surface protein that is expressed on various cell types, including T cells, B cells, and monocytes. The antibody is conjugated with the fluorescent dye APC, which can be used for flow cytometric analysis.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd9 pe, by BioLegend (1 mentions)
Anti cd63 pe, by BioLegend (1 mentions)
Aldehyde sulfate latex beads, by Thermo Fisher Scientific (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Anti cd19 alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Facsdiva software, by BD (1 mentions)
Facs fortessa, by BD (1 mentions)
Anti cd63 apc, by BioLegend (1 mentions)
0.2 μm filter, by Merck Group (1 mentions)
Memglow 488, by Cytoskeleton (1 mentions)
4 protocols using anti cd81 apc
1
Comprehensive Exosome Characterization
2
Characterization of CD19 and CD81 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293T cells (RRID CVCL_0063) were seeded at 100,000 cells/well in 24 well plates 12–18 hours prior to transfection. Cells were transfected using Lipofectamine 2000 with either with either 1.5 μg of empty pcDNA3.1(+) vector, 0.75 μg of CD19 DNA and 0.75 μg of empty pcDNA3.1(+) vector DNA (CD19 condition), or 0.75 μg of CD19 DNA and 0.75 μg of CD81 DNA (CD19+CD81 condition). 36–48 hours after transfection, cells were harvested in phosphate buffered saline (PBS) supplemented with 3 mM EDTA, transferred to a 96 well V-bottom plate, and then washed twice with PBS. Cells were then incubated on ice for 20 minutes with 2 μg/mL anti-CD19-Alexa Fluor 488 (Thermo Fisher) and anti-CD81-APC (BioLegend, Catalog number 349510, RRID AB_2564021) in 20 mM HEPES buffer pH 7.4, containing 150 mM NaCl, and 0.1% BSA. Cells were washed two times with PBS and analyzed on a Beckman Coulter CytoFLEX.
3
Flow Cytometric Analysis of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The flow cytometry method was applied to determine the expression of CD63, CD9, and CD81 on the surface of isolated particles. Isolated TEx or TMv were resuspended in PBS filtered through 0.2 μm filters (Merck Millipore) and then labeled with monoclonal antibodies conjugated with fluorochromes: anti-CD63 APC, anti-CD9 APC, anti-CD81 APC, rat IgG2aκ APC and Armenian hamster IgG APC isotype controls (all from BioLegend). The expression of cell surface markers was analyzed using FACS Fortessa with FACSDiva software (Becton Dickinson).
4
Single-EV Analysis by Imaging Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 108 EVs were analyzed at the single EV level by imaging flow cytometry (ImageStream X MKII, Amnis/Luminex) following guidelines and principles described in Görgens et al. (28 (link)) (see SI Appendix ), with the following labels: lipid dye MemGlow 488 (Cytoskeleton, Inc), anti-CD81-APC (dilution 1/25) (Clone 5A6, Biolegend), and anti-CSF-1-PE (dilution 1/25) (Clone 26786, R&D Systems). Adequate PBS, unstained, Membright488, CD81, CSF-1 monocolors, and fluorescence minus one (FMOs) EVs controls were performed in parallel.
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!