Multiplex Luminex® Assays
Multiplex Luminex® assays enable the simultaneous analysis of multiple proteins in single samples from a broad range of biological sources. Based on Luminex® xMAP® (multi-analyte profiling) technology, Invitrogen™ multiplex assays combine the efficiency of multiplexing with the accuracy, sensitivity, reproducibility, and simplicity of ELISA. We make Luminex® xMAP® technology simple and reliable to use by providing both nonmagnetic (polystyrene) and magnetic kits, which include ready-to-use validated combinations of bead-coated and detector antibodies, microplates and other reagents.
Features of Invitrogen multiplex kits for Luminex® assays
Our multiplex kits undergo rigorous quality testing and are calibrated against matching Invitrogen™ ELISA kits where available (Table 1) to show comparable analytical results between protein analysis platforms.
- Fast and efficient—simultaneous analysis of up to 100 proteins using only 50 μL
- Accessible—a broad menu of Invitrogen immunoassay kits based on xMAP® technology
- Economical—helps reduce time and costs compared to running multiple western blots and ELISA assays
- High specificity—lower cross-reactivity with other targets (Table 2)
Figure 1. Strong correlation of ELISA and Invitrogen multiplex assay results. Mouse GM-CSF in tissue culture supernatant was tested by ELISA and in an Invitrogen™ (Novex™) multiplex assay for Luminex platform. Correlation of values over 3 orders of magnitude of sample dilution was 0.9868.
Table 1. Rigorous assay validation of Invitrogen multiplex kits helps ensure consistent, reliable results.
|Benchmarking to ELISA
|Correlates with 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 (Figure 2)||Inter-assay CV: <10%
Intra-assay CV: <10%
|Specificity||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 (Figure 3)||(R2 > 0.99) Recombinant standards are compared to natural samples to ensure equivalency|
Figure 2. Assay precision. Orange and gray bars each represent 24 replicates measured on separate days. CVs in all cases were <10%. Data were generated using a Invitrogen™ Human Cytokine Magnetic 10-Plex Panel (Cat. No. LHC0001M).
Figure 3. Parallelism to natural samples. To evaluate Invitrogen™ (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 Invitrogen multiplex assay kit and using similar kits from two other suppliers. The multiplex sample was quantified on the MAGPIX® system, and percent recovery calculated for (A) the individual markers and (B) as an average for the entire group.
Table 2. Specificity data from the Human Cytokine 10-Plex Panel (Cat. No. LHC0001) show no significant cross-reactivity.
Numbers represent mean fluorescence intensity (MFI) units generated when individual recombinant proteins were analyzed independently in a series of 10-plex assays.
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