GeniePlex FAQs

GeniePlex assays can be run on most flow cytometers equipped with a single blue, or both blue and red lasers. The maximum emission of the bead classification dye is 700 nm. It can be detected on PE-Cy5 channels of most of the flow cytometers with blue (488 nm) excitation. It can also be detected on PE-Cy7 channels with blue (488 nm) excitation or APC channels with red (633 or 640 nm) excitation if such a fluorescence channel is available. See below.

NOTE 1: Both areas and heights of the FL parameters should be collected

NOTE 2: *Trademarks are the property of their respective owners

For FC500, Gallios and Navios flow cytometers from Beckman Coulter, blue and red lasers are collinear. For this reason, colour compensation is usually needed between PE and APC.

The Attune NXT flow cytometer, if equipped with a blue and a red laser, can acquire PE (assay reporter) and APC (bead classification) signals. However, fcs files exported from this system cannot be analyzed using FCAP Array v3. Moreover, GeniePlex assays have not been tested on this system to date. Therefore, the sensitivity of the NXT flow cytometer for GeniePlex assays is unknown.

Guava is not a flow cytometer. It does not use hydrodynamic focusing and does not have good sensitivity. We do not recommend it for GeniePlex assays.

We have a selection of 400 analytes for which multiplex panels can be created for. These panels can be created in as little as 20 days. To find out more information about developing your own custom panel, please click here to get in touch.

The reconstitution and preparation of serial dilutions of the antigen standards should be done no more than 2 hours before carrying out the assay(s). Diluted standards should be discarded after use. Freezing, thawing and reuse of the re-constituted and diluted standards are NOT recommended.

If for some reason the assayed plate cannot be read immediately, it can be stored at 2-8°C for up to 16 hours. In this case, the plate should be sealed with a plate seal and protected from light by wrapping the filter plate in aluminum foil until used.

We recommend 3-fold dilution for serum, plasma, cell culture supernatant, and tissue lysate samples. With the exception of tissue lysate samples, normalization is usually not needed for samples. If normalization for tissue lysate samples is required, the Lowry Protein Assay is recommended to establish protein concentration.

While we do not have any citations describing the use of GeniePlex with CSF samples, we do have partners using it with tissues and serum/plasma. Normal CSF samples have similar pH and buffer conditions as serum plasma. In many cases, this sample type contains much less proteins and no platelets or cells (due to the compartmentalization effect of the blood-brain barrier). Samples should be centrifuged to remove cells and/or platelets possibly present in the sample. It is recommended to use the SPB (Serum/Plasma/Bodily Fluid) Diluent Kit when preparing CSF samples since CSF has a less complex matrix.

We have not extensively tested GeniePlex with urine samples. However, there is experimental evidence showing 2 cytokines can be detected simultaneously in urine samples with GeniePlex.

In general, the majority of GeniePlex assays acquired on most flow cytometers have quantitation ranges of < 20 pg/mL to > 5,000 pg/mL. Therefore, there should be less chance of needing to repeat an assay for issues associated with range.

Yes. Customers can use FlowJo to identify bead populations and obtain PE Mean Fluorescent Intensies (MFIs) of each bead set. For concentration calculations, users can use a commercially available software package.  

GeniePlex is an excellent alternative to standard ELISA technology for a number of reasons. GeniePlex enables the simultaneous and quantitative detection of up to twenty-four analytes in as little as 15μl sample. For the same amount of analytes to be tested by ELISA, 50-700µL of sample would be needed, run on 7 different sample plates. GeniePlex is also a quicker procedure to carry out, taking 4 hours. ELISAs can take up to 7 hours to complete. Moreover, the assay range of GeniePlex assay kits is 2-5000pg/mL, with the range of ELISA kits only 2-1500pg/mL.

GeniePlex gives excellent performance versus Luminex technology. GeniePlex Human TH1/TH2/TH17 7-Plex assay and a Luminex multiplex assay were used to assay cell culture supernatant samples from human PBMCs treated with various reagents. Results showed a strong correlation of GeniePlex Multiplex Immunoassays with Luminex xMAP technology. See figure 4 of https://www.assaygenie.com/multiplex-elisa for study findings.