How xMAP® Technology Works

Luminex® xMAP® technology enables scientists to measure multiple proteins in a single well. This technology combines advanced fluidics, optics, and digital signal processing with proprietary microsphere technology to deliver multiplexed assay capabilities. Featuring a flexible open-architecture design, xMAP® technology can be configured to perform a wide variety of bioassays quickly, cost-effectively and accurately.

Microsphere beads, either polystyrene or paramagnetic, are color-coded into up to 500 distinct sets. Each bead set can be coated with a reagent specific to a particular bioassay, allowing the capture and detection of specific analytes from a sample. Inside the Luminex® analyzer, a light source excites the internal dyes that identify each microsphere particle, and also any reporter dye captured during the assay. Multiple readings are made on each bead set, which further validates the results. Using this process, xMAP® technology allows multiplexing of up to 500 unique bioassays within a single sample, both rapidly and precisely. xMAP® technology is compatible with the following Luminex® analyzers:

  • MAGPIX® system—affordable, efficient, and compact
  • Luminex® 100/200™ system—versatile, efficient, and widely used in multiplexing
  • FLEXMAP 3D® system—high throughput, up to 500 simultaneous assays, and automation compatible

Novex® multiplex immunoassay tips and tricks

Multiplex Kits

Our multiplex kits undergo rigorous quality testing (Table 1) and are calibrated against matching Novex® ELISA
kits, if available, so that both protein analysis platforms will provide comparable analytical results. Novex®
multiplex kits are:

  • Fast and efficient—simultaneous analysis of multiple proteins using only 50 μL of precious sample
  • Accessible—broad and expanding menu of Novex® immunoassay kits based on xMAP® technology
  • Economical—can significantly reduce time and costs compared to running multiple western blots or ELISA assays

Life Technologies makes the use of Luminex® xMAP® technology simple and reliable by providing a range of Novex® singleplex and multiplex immunoassay kits, a full line of multiplexing Luminex® instruments, xPONENT® data analysis software, and extensive assay and instrument technical support, including on-site demonstrations and personalized technical consultation services.

Specification Description
Benchmarking to ELISA (see Figure 1) Correlates to ELISA data (>90% correlation)
Recovery Tested on serum and plasma
Sensitivity Physiologically relevant levels, <10 pg/mL (based on detectable signal >2 SD above background)
Precision (see Figure 2)
  • Inter-assay CV: <10%
  • Intra-assay CV: <10%
Specificity (see Table 2) Cross-reactivity tests are performed with other analytes and antibodies
Linearity of dilution High coefficient of correlation between sample dilutions and expected concentration over the range
of the assay
Parallelism to natural samples (see
Figure 3)
(R2 > 0.99) Recombinant standards are compared to natural samples to ensure equivalency


Table 1. Rigorous assay validation of Novex® multiplex kits helps ensure consistent, reliable results.


How to prepare your Novex® multiplex assay


Figure 1. Multiplex assay results correlate to ELISA assay data. Murine GM-CSF in tissue culture supernatant fluid was tested by ELISA and in a Novex® multiplex assay. Correlation of values over 3 logs of sample dilution was 0.9868.


Figure 2. Assay precision. Green and gray bars each represent 24 replicates measured on separate days. CVs in all cases were less than 10%. Data were generated using a Novex® human cytokine 10-plex magnetic assay kit.

How to analyze multiplexed protein data


Figure 3. Parallelism of recombinant standards to natural samples. To evaluate the Novex® multiplex assays, samples of 23 human protein markers were spiked into a sample of human serum, and the sample was processed using the manufacturer’s instructions for the Novex® multiplex assay kit and using a similar kit from two other suppliers. The multiplex sample was quantified on the MAGPIX® system, and percent recovery calculated (A) for the individual markers and (B) as an average for the entire group.


Antibody Bead Kit

  IL-1ϐ IL-2 IL-4 IL-5 IL-6 IL-8 IL-10 TNF-α IFN-γ GM-CSF
IL-1ϐ 6,332 41 37 13 9 37 32 27 18 51
IL-2 38 11,324 47 17 26 40 33 27 15 50
IL-4 38 41 10,503 15 25 40 32 25 12 50
IL-5 48 54 36 8,265 11 34 33 25 16 51
IL-6 44 72 40 11 6,827 42 31 22 12 70
IL-8 39 39 44 12 27 10,002 32 27 19 52
IL-10 41 51 39 11 12 41 8,151 23 12 46
TNF-α 32 55 36 10 7 60 30 6,642 10 47
IFN-γ 32 41 39 10 8 70 35 25 3,540 51
GM-CSF 41 40 36 12 8 46 33 25 13 4,932


Table 2. Specificity data from the human cytokine 10-plex panel shows no measurable cross-reactivity. Numbers represent mean fluorescence intensity (MFI) units generated when individual recombinant proteins were analyzed independently in a series of 10-plex assays.