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The Quansys Multiplexed ELISA Array consists of a non-contact printed array of capture antibodies to a variety of either mouse or human cytokines. The user adds
an antigen standard and samples to individual wells of a 96-well plate. After incubation and washing, a mixture of biotinylated antibodies is added to the plate.
This is followed by incubating with streptavidin-HRP and is completed by capturing the light produced from adding chemiluminescent substrate with a CCD imaging
system. The chemiluminescent signals from individual spots are measured using a CCD camera. Pixel intensity values are used to construct a standard curve individual
standard curve and calculate the concentrations of the unknown sample for each system.
Yes, the array typically uses a five-point standard curve to accurately quantify samples. The five-point curve has been designed to encompass the biological ranges
for individual cytokines.
No, in some cases the typical five-point standard curve does not reach the LLD for a given cytokine. The user has an option of using additional dilutions of the
antigen mix to achieve the LLD for a given cytokine.
We recommend testing each sample in step dilutions. For example, when testing cell culture supernatant for unknown cytokine concentrations; a 1:2, a 1:10 , and
a 1:50 dilution are typically used.
The standard curves shown in the Certificate of Analysis use pixel intensities on the Y-axis against concentrations on the X-axis. Will different
CCD cameras give different pixel intensities and how will this affect my assay? Yes, different CCD cameras give different pixel intensities based on the sensitivity
of the camera. However, this will not affect assay performance or standard curve generation as the pixel intensities are relative values.
Once the lyophilized reagents are reconstituted they should be used immediately. If the user desires to run several assays using the 96 well plate, the reconstituted
antigen standards should be immediately aliquoted and frozen at -20°C. The entire kit contents should be used within one week of reconstitution. Detailed information
on kit storage after reconstitution can be found in the User's manual.
No, for best results it is recommended that the assay be completed once started. After reading the results with a CCD camera, the user has the option to delay
data analysis to a convenient time.
The user will need to provide a pipette and CCD imager. We also recommend an automatic plate washer for best results.
The Quansys Multiplexed ELISA Array plate can be read by a number of CCD imaging systems and gel documentation systems such as Alpha Innotech Fluorchem SP, Kodak
4000 MM, Fujifilm LAS-3000, Bio-Rad Versa Doc 4000, and others. A cooled and non-cooled CCD imager with at least 12-bit outputs contained in a light tight box
are appropriate for imaging. Our array format and the Quansys Array Software are compatible with most CCD imaging systems. High-resolution cameras in light tight
boxes with at least 19" from the lens to the plate if imaging from the top or 6" from the bottom are ideal for this application, but not required.
The software allows the user to open the plate image, mark the spots, and analyze the pixel intensity of individual spots on the plate. This software does not
automatically find spots or provide detailed data reports (standard curve plots or statistical analysis). Data should be exported into Microsoft® Excel®
or a similar program for further analysis and data management.
The Quansys Multiplexed ELISA Array has been tested with serum, plasma, whole blood, urine, and cell and tissue lysate. We have experienced no interference from
FBS, 1% NP-40, 1% Tween, 1% Triton X-100, heparin (30 mg/ml), 1M urea, 20 mM EDTA, 20% citrate. SDS has been shown to interfere with the assay. Human samples
containing human antibodies must be diluted 1:2 in the provided sample dilution buffer. If tissue homogenates or cell lysates are used, centrifuging is recommended
to clarify samples prior to using in the assay. When using tissue homogenates, we recommend preparation in a protein-free buffer so that samples can be tested
for total protein to normalize cytokine responses.
The Quansys Q-Plex™ Array requires 30 µl total volume per well
(sample plus diluent). Smaller sample volumes may be used when high levels
of bio markerss are anticipated. Mouse samples can be run neatly. However,
human samples containing human antibodies MUST be diluted 1:2 in sample
dilution buffer prior to running the assay.
Most protocols require as little as two and a half hours for completion. After imaging the plate, data analysis may be carried out later.
The Quansys Multiplexed ELISA assay has <15% CV between kit lots and has been validated at <25% error when compared to known controls.
Yes, Quansys Biosciences specializes in assay development, please call or email for details and pricing.
Yes, your samples can be mailed to Quansys where they will be tested using the Quansys Multiplexed ELISA platform. Data, along with statistical analyses will
be provided to the user within two weeks. Please call for shipping details and pricing.
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