Overview of ELISA Technology
Here we present an overview of ELISA technology, some of the basic ELISA procedures, the Life Technologies products best for your unique situation, some specific references, and, finally, tips and tricks from our scientists that will speed your success.
Life Technologies offers two options for performing ELISAs: antibody pairs and ready-to-use ELISA kits (Table 1). Novex® Antibody Pairs are matched pairs of detection and capture antibodies for researchers who prefer to coat their own ELISA plates or wish to use another platform for protein analysis. Novex® ELISA kits are complete, ready-to-use kits with precoated plates, buffers, and capture antibodies included.
|Novex® Antibody Pairs||Novex® ELISA kits|
|Ready-to-use reagents?||No; need overnight coating process||Yes|
|Total assay time||Overnight plus 4 hr||2.5–4 hr|
|Readout||HRP-TMB (colorimetric)||Varies (colorimetric or chemiluminescent)|
|Instrumentation needed||Microplate reader||Varies (absorbance or luminescence microplate reader)|
Learn how to use Novex® Antibody Pairs to set up an ELISA, and find out what pre-matched antibody pairs we offer to help make ELISAs easier and more successful. Visit Novex® Protein Handbook Registration Form to learn more.
Learn all about running an ELISA to measure target proteins in serum, plasma, supernatants, lysates, and other sample types. The ELISA is a widely accepted method for quantifying selected proteins and is often used in conjunction with western blot to analyze proteins in research samples. Visit Novex® Protein Handbook Registration Form to learn more.
- Novex® ELISA kits can be further categorized into several different groups (Table 2), based on a number of factors: target protein class, sensitivity, readout method, or ability to detect specific phosphorylation states of the target protein.
- Novex® phosphoELISA™ kits enable the specific detection of phosphorylation of key signaling proteins with high specificity, and are often used to supplement western blot data and provide quantitative data.
- Novex® Chemi ELISA kits enable protein quantitation across a wide range of sample concentrations, minimizing guesswork for sample dilutions.
- Novex® Ultrasensitive ELISA kits use a standard colorimetric readout but enable detection and analysis of proteins to levels as low as 0.5 pg/mL, especially useful with highly diluted samples.
|Standard colorimetric ELISA||<1 pg/mL||<1 pg/mL||<1 unit/mL|
|Analytical sensitivity*||<10 pg/mL||Yes|
|Measurement range*||5–250 pg/mL||0.5–20 pg/mL||0.5–2,000 pg/mL||1.6-100 units/mL|
|Incubation time*||4 hr||4–5 hr||3.5 hr||4 hr|
|Readout||HRP-TMB (colorimetric)||HRP-TMB (colorimetric)||AP-CSPD
|Instrumentation needed||Microplate reader||Microplate reader||Microplate reader
capable of measuring
|Able to detect
*Every assay has its own unique specifications. Please consult the manual associated with your specific Novex® ELISA kit.