vTAG™ ANTI-HUMAN CD63 ANTIBODY
NO WASH, QUANTITATIVE CD63 MEASUREMENT BY vFC™
$105.00 – $275.00
65 in stock
Est. Limit of Detection: 30 Molecules per EV
Cargo analysis using vTag™ antibodies should be accompanied by a vesicle specific analysis method such as vFC™. vTag™ antibodies bundled with the vFC™ assay will be discounted 25%.
Certificate of Analysis
CD63 Immunostaining in vFC™?
This protocol provides a standardized means of analyzing individual vesicles to measure size, concentration, and CD63 expression simultaneously via vFC™. vFC™ (#CBS4) is an assay that enables specific detection and characterization of individual extracellular vesicles such as exosomes and microvesicles on commercially available flow cytometers. Because vFC™ is vesicle specific, we can characterize vesicles and cargo in a highly-reproducible, no-wash assay without artifacts from debris or reagent aggregates. In this protocol, vesicles will be stained using our vTag™ Anti-Human CD63 Antibody. See protocol 2 in the vFC™ kit handbook for additional considerations and full protocol details.
- Dilute sample containing EVs to approximately 5×106 extracellular vesicles per µl. To determine vesicle concentration for an unknown sample, see supporting protocols included in the vFC™ kit.
- Prepare a sufficient amount of vFluor™ Red for all samples and controls (5 µl per well) according to kit instructions
- In duplicate, in the included 96 well plate, add 35 µl of dilution buffer, 5µl of prepared vFluor™ Red, 5 µl of vTag™ anti-human CD63 antibody, and 5 µl of EV samples.
- Incubate in the dark at room temperature for 1 hr.
- Perform serial dilution of the samples as described in the assay handbook. Briefly, samples are to be diluted from between 1:100 to 1:1000 from staining concentration depending upon the flow cytometer being used.
- Run the diluted sample of labeled EVs on a qualified flow cytometer.
CD63 is a member of the tetraspanin family. In cells, tetraspanins are important to many biological functions including cell adhesion and motility as well as signaling and protein trafficking.
CD63 predominantly associates with the outer membrane of MVBs in cells leading researchers to originally describe it as a universal marker for vesicles originating within cells, principally exosomes. Recent evidence, of course, suggests that there are populations of exosomes and other vesicles that do not express measurable amounts of CD63. More information on CD63 coming soon!
20 Tests, 100 Tests