TRIFicTM assays are capable of detecting small amounts of exosome bound antigens, even in unpurified samples
Antibodies against the CD antigens are available. These have all been validated for western blot analysis
ExoFLARETM tracking system provides unsurpassed sensitivity for tracking exosome markers.
TRIFic™ exosome assay kit
- EX101, CD9 TRIFic™ exosome assay
- EX102, CD63 TRIFic™ exosome assay
- EX103, CD81 TRIFic™ exosome assay
ExoFLARE™ exosome tracking system
- EX301, ExoFLARE CD9 Expression construct, 20 ug
- EX302, ExoFLARE CD63 Expression construct, 20 ug
- EX303, ExoFLARE CD81 Expression construct, 20 ug
- EX401-100, EF Red xc cell impermeable dye, 100 reactions
- EX401-1000, EF Red xc cell impermeable dye, 1000 reactions
- EX402-100, EF Red s cell permeable dye, 100 reactions
- EX402-1000, EF Red s cell permeable dye, 1000 reactions
- EX205, Trial Kit containing: CD9 Clone CGS12A, CD63 Clone CGS82X and CD81 Clone CGS36K, 3 * 20 ug
- EX201-100, CD9 Clone CGS12A monoclonal antibody, 100 ug
- EX202-100, CD63 Clone CGS73B monoclonal antibody, 100 ug
- EX204-100, CD63 Clone CGS82X monoclonal antibody, 100 ug
- EX203-100, CD81 Clone CGS36K monoclonal antibody, 100 ug
The TRIFic™ exosome assay is similar to an ELISA assay however there are some significant differences. Unlike an ELISA assay, there is no enzymatic reaction. Rather, the target is detected directly with a Europium label. TRIFic™ exosome assays quickly provide quantitative data from purified or unpurified samples, including direct measurement of exosomes in plasma in a convenient 96-well format. TRIFic™ exosome assays are being made available for widely used markers of exosomes; the tetraspanin proteins CD9, CD63 and CD81. TRIFic™ exosome assays deliver clear, consistent data.
• Simplicity of the assay results in a high degree of reproducibility.
• Europium is used for detection, providing a high degree of sensitivity.
• Only antigens displayed in multiple copies on a support within the sample (as with tetraspanins on exosomes) are detectable with the assay, resulting in increased specificity
ExoFLARE™ constructs were transiently transfected into DU154N cells. ExoFLARE™ cell-permeable dye was added to the media and cells were imaged using a confocal fluorescence microscope.
Exosome antigen antibodies from Cell Guidance Systems detect some of the most widely studied exosome antigens. The antibodies have been validated for the detection of exosomes and offered the same antibodies used in the TRIFic™ assay kit.
In the TRIFic™ exosome assay, the same antibody is used for binding of target to the assay plate and for detection. The assay consists of a monoclonal antibody (labeled with biotin) bound to a streptavidin coated plate which captures protein present in the surface of exosomes. Subsequently, an identical monoclonal antibody (labeled with Europium) is used for detection. Because the capture and detection antibody are identical, they require two linked copies of the same epitope for a signal to be detected. Exosomes provide an ideal structure to link CD9 molecules and allow detection of CD9 in this assay. Exosomes typically have multiple copies of CD9 facing towards the attachment surface and additional CD9 molecules available for detection. Any non specific binding of capture and detection antibodies is unlikely to generate a signal. Using a Europium fluorophore provides high levels of sensitivity for the assay, which is able to detect small changes in the abundance of the target CD9 protein even within unpurified complex biological samples, such as blood plasma and cerebrospinal fluid.
Principle of ExoFLARE™ detection. ExoFLAREs™ utilize a combination of a FLARE (FLuorescence Activating Response Element) protein tag together with a pro-fluorophore dye. Neither the protein or dye fluoresce in isolation. When the protein binds to the dye, it causes a change in structure which results in fluorescence. The dye and protein form an unstable bond with continuous turnover of the dye. This results in sustained fluorescence without the levels of photo-bleaching associated with fluorescent proteins. This allows ExoFLAREs™ to be monitored for extensive periods to allow tracking of movement of dyes.
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