Overview of Multiplex Immunoassays

Traditionally, single-analyte, or singleplex, protein detection methods such as enzyme-linked immunosorbent assays (ELISA) or western blotting have been used sequentially to analyze multiple intracellular and extracellular proteins. Although these are both well-established, validated techniques, they can be time-consuming, sample-depleting, and costly when used to measure numerous markers per sample. Life Technologies offers a range of Novex® assays on the Luminex® xMAP® (multi-analyte profiling) technology platform for detection and quantitation of multiple proteins simultaneously, saving time, sample, and money.

  • Efficient—simultaneously analyze multiple proteins using only 50 μL of sample
  • Economical—helps save time and costs compared to western blot or ELISA
  • Simple—easy operation with streamlined start-up and shutdown protocols

Order Luminex® Protein Assays

Novex® multiplex immunoassays enable the simultaneous analysis of multiple proteins in single samples from a broad range of biological sources. They combine the efficiencies of multiplexing with the accuracy, sensitivity, reproducibility, and simplicity of ELISA.

 

Measure multiple proteins simultaneously using the Novex® multiplex bead-based kit on the Luminex® instrument platform
  Figure 1   Figure 2
elisa procedure
 
Figure 1. Fast and easy, 4-hour Novex® ELISA kit protocol. Capture antibodies are precoated on the bottom of a 96-well plate. The sample or standard is added to the wells and incubated to allow target proteins to bind. The wells are then washed to remove unbound material, and the detection antibody is added and incubated to form a sandwich around the protein of interest. HRP conjugate is then added and incubated to allow binding via a biotin–streptavidin interaction. Next, the chromogen substrate for HRP is added, and the subsequent enzymatic reaction turns the solution blue. Finally, the reaction is stopped, turning the solution yellow in proportion to the amount of target protein in the sample. Absorbance is read in a microplate reader at 450 nm.
 
Figure 2. General schematic of phosphoELISA™ protocol. Capture antibodies are precoated on the bottom of a 96-well plate. The cell extract or standard is added to the wells and incubated to allow target proteins to bind. The wells are then washed to remove unbound material, and the detection antibody is added and incubated to form a sandwich around the protein of interest. HRP anti-rabbit antibody is then added and incubated to allow binding to the rabbit-derived detection antibody. Next, the chromogen substrate for HRP is added, and the subsequent enzymatic reaction turns the solution blue. Finally, the reaction is stopped, turning the solution yellow in proportion to the amount of target protein in the sample. Absorbance is read in a microplate reader at 450 nm.