EVix Multi-Tope ELISA Kit 256 Wells/115 Ab
Research Use Only Not for Diagnostic Use
Description: EVix Multi-Tope ELISA kit brings revolutionary screening capabilities to EV assays. Using highly specific monoclonal antibody, captured EVs are detected with a detection antibody mixture specific to CD9, CD63, and CD81. Expression of multiple tetraspan molecules on single EVs, allows even small numbers of captured EVs to be detected and quantified.
The Multi-Tope ELISA kit contains duplicate wells for each capture antibody, one to capture EVs and the other is a negative control. The signals generated are specific for EV captured with the unique capture antibody within the individual wells.
The power of the Multi-Tope assay is its unique capture plates which include 115 unique capture antibodies. This complete assay kit allows assessment of 115 unique antigens in an EV sample.
All capture antibodies are certified and coated at optimum concentrations to yield optimal signals. The capture antibodies are selected from our catalogue of surface and immune associated antigen specific monoclonal antibodies. Each antibody has been tested and qualified by flow cytometry and additional information is provided by the manufacturer for each clone. Reference Ancell catalogue for information on each clone including performance data sheet of each antibody's performance.
Production of each Multi-Tope ELISA kit is done under rigorous controls to assure consistent performance from lot to lot. All antibodies are qualified by flow cytometry and tested for sensitivity and specificity. Coating of the individual wells is done using high precision pipetting.
The Multi-Tope ELISA kit comes complete with all reagents needed to run the 256 well assay. To assure maximum signal, an overnight capture is followed by a 1 hr detection incubation. With minimal effort, an EV preparation can be finger printed for the presence of 115 unique antigens associated with expression of CD9,CD63, and CD81.
Multi-Tope Plate Maps
Each capture antibody is mapped for the Multi-Tope plates provided in the kit.
Paired wells provide the negative control well next to the experimental. Odd columns do not receive antigen and provide a non specific background signal which is subtracted from the experimental well containing EV preparation.
Each capture antibody has a performance data sheet available as described above. By going to the Ancell Catalogue and searching the provided catalogue number, a pdf of the antibody performance data is available.
The Multi-Tope assay kit comes complete with capture plates and all reagents needed to complete the assay. An overnight capture followed by a 1 hour detection incubation yields a high definition fingerprint of the antigen profile of your EV preparation.
Use to compare and contrast EV preparations from different sources, conditions, or production runs. Identify unique antigens associated with your EV samples.
Finger Print your EV preparations.
EVix Multi-Tope assay brings revolutionary screening power to EV biology. Each assay screens for over 100 unique cell surface antignes. If the antigen is present, the EV is captured. The detection antibody cocktail of anti CD9, CD63, and CD81 antibodies mazimizes signal and specific antigen bearing EVs are detected.
Unlike Mass Spectrometry, the Multi-Tope assay reads only membrane/tetraspan associated signals captured by specific antigen. Hundreds of antigens are reportedly EV associated, however many of these studies run entire preparations of EVs that have been concentrated by centrifugation or precipitation and co-seperated molecules can lead to detection of molecules not necessarily associated with membranous vessicles.
The Multi-Tope assay screens your EV preparations quickly and thoroughly creating a unique phenotype. Much like a finger print, the Multi-Tope assay can be used to confirm similarities or differences between EV preparations.
Screen for Antigen Presence.
EVix Multi-Tope assay brings revolutionary screening power to EV biology. 13 ml of EV preparation adjusted to give a HSTT value of 3 or higher yield antigen profiles specific to the preparation.
Negative control signals are also shown, which are background signals without EV sample. This sample is normal human urine, concentrated 100X by diafiltration against a 300,000 MWCO membrane. Samples were dia filtered against TBS glycine solution. Note presence of EPCAM (CD326) on tetraspan specific EVs obtained from normal unringe. Neprilysin (CD10) is also present on urine EVs.
Phenotype EV Preparations.
Antigen signals vary significantly between EV origin. The panel on the left shows results from a human mixed lymphocyte cultrue supernatant harvested on Day 7 of culture.
In contrast to the urine antigen patterns, the EV phenotype obtained from a mixed lymphocyte culture sample shows the presence of more immune associated molecules.
Negative control signals are not shown but have been subtracted out from the positive values. This sample is culture supernatant that has been concentrated by diafiltration in TBS glycine against a 300,000 MWCO membrane.
Rank Antigen Signals.
Antigen signals not only vary between different antigen preparations, but also within preparations. Since the antigen signal is quantitative, the antigen specific signals can be ranked and plotted according to intensity. As shown left, the signals from pooled culture supernatants run at 50X concentration have a hierarchy and can be used to further characterize EV populations.
Negative control signals are not shown having been subtracted out from the positive values. This sample is a pool of different cell culture supernatants that has been concentrated by diafiltration in TBS glycine against a 300,000 MWCO membrane.
Compare and Contrast EV Populations.
Shown left is an XY scatter plot of antigen signals for a Urine sample (Horizontal) and mixed culture supernatant (Vertical) obtained by Multi-Tope assay.
While the antigens in the upper right corner appear to be shared in both preparations, there are distinct antigens present only in one of the preparations, and fall close to the axis. The antigens shared are tetraspan like and integrin related. Preparation unique antigens are certain cell surface antigens.
Look for more than 115 surface antigens in a single assay
Screen cells, membrane preps, EVs for presence of 115 antigenic epitopes.
Assay multiple samples with high sensitivity and reproducible results.
Compare transcripts to actual protein
Screen transcripts and compare to actual protein presence on membrane surfaces
A simple assay yields both confirmatory but also quantitative values for the presence of 115 antigens.
Compare differences populations
Quantify unique antigen profiles associated between different samples and clinical conditions. The absence or presence of antigens can be distinctive and related to sources and conditions. Patterns can be tracked quickly to identify target molecule differences.
The Surveyix plates preserves 115 antibodies in individual wells which can be used in a single assay. Multiple plates can be easily run to accommodate more samples or experimental conditions.
Test entire atlas of immune associated surface molecules
The current SurVeyix plates contain a collection of immune associated antibodies which detect epitopes expressed on the cell surface and can also be detected on EVs and membrane preparations.
Target protein/peptide epitopes
Unlike transcripts the SurVeyix plates allow for detection of functioning epitopes unique to each antibody. In conjunction with transcript analysis, powerful screening of cellular responses and different populations is possible.
A: Multi Tope Capture Plates: Three (MT1, MT2 and MT3) 96 well plates (Store 4C) containing 256 individual assay wells
B: 10X Sample and Wash Buffer Concentrate: 1 bottle (40 ml) to make 400 ml. (Store 4C)
C: Biotinylated Primary Detection Antibody: 1 vial of 1.5 ml to make 30 ml. (Store -70C)
D: Horse Radish Peroxidase Conjugated Streptavidin(SAHRP): 1 vial of 1.5 ml to make 30 ml. (Store -70C)
E: Chromogenic Solutions : 1 bottle (30 ml) of a stabilized H2O2 , 1 vial (1.5 ml) of stabilized TMB chromogen (Store 4C) Caution!
F: Stop Solution: 1 bottle (30 ml) Acid Caution! (Store RT)
If contents are unopened and stored at 4C and -70C the performance of the kit is stable for 6 months.
Order MT-001. Complete kit for 256 well assay.
Catalog MT 001