6 ELISA test formats for protein target and biomarker discoveries


Having access to highly reproducible and fully validated ELISA tests is crucial in today’s Life Science research. Although the Sandwich ELISA format is the most commonly used, other immunoassay formats are also available to allow for a complete tool box, from phosphorylation studies to transcription factor activity assays.

In this post, I’ll be taking a look at the 6 formats available.

Sandwich ELISA test principle

Sandwich ELISA test principle

For example, RayBiotech (distributed in Europe via tebu-bio) manufactures over 2 600 ISO certified and rigorously tested ELISA tests, all backed by a 100% guarantee. All RayBio® ELISAs are validated for exceptional reproducibility. Every kit is subjected to a battery of rigorous product-specific quality control tests and manufactured in compliance with GMP and ISO 13485 standards.

Currently 5 formats are available:

#1- Sandwich ELISA

This matched antibody pair based format allows for the detection of cytokines, growth factors, soluble receptors and other soluble proteins. Samples such as plasma, serum, culture media, lysates or clarified body fluids are suitable. A wide variety of species are available such as human, rat, mouse but also cow, dog, horse, cat, sheep, pig, rabbit, rhesus macaque and cotton rat!

Reproducibility and Precision

These sandwich ELISA kits are evaluated for intra-assay reproducibility by running 2-3 positive control samples in duplicate on a single plate (maximum tolerance = 10% CV). Inter-assay reproducibility is evaluated by at least 2 independent experiments with 2-3 positive control samples and a full standard curve (maximum tolerance = 12% CV).

Lot-to-lot consistency is tested by comparing calculated concentrations of 2-3 positive control samples with calibration curves of current lot and previous lot (maximum tolerance = 20% CV).

Dilution Linearity and Recovery

Recovery is determined by spiking various levels of target protein into biological samples. Dilution linearity is tested by performing 2-fold and 4-fold dilutions of biological samples. Recoveries typically range from 80-130% (maximum tolerance 70-150%). Tested biological sample types include serum, EDTA plasma, citrate plasma, heparin plasma (normal healthy donors) and cell culture medium (DMEM or 1640). Recommended dilution ranges for serum and plasma are determined from this testing series. For lysate-specific sandwich ELISAs (catalog numbers ending in “-CL”), sample types include cell lysates and tissue homogenates of various origins.

Access to our selection of Sandwich ELISA Kits here!


Phosphorylation ELISA test

Phosphorylation ELISA test

#2– Phosphorylation ELISA

Also sandwich based, these assays can detect phosphorylated targets such as MAPK, Akt, EGFR, Stat, mTOR and other signaling molecules. They are compatible with cell and tissue lysates for human, mouse and rat proteins.


Access to our selection of Phosphorylation ELISA Kits here!




#3- Cell-Based-phospho-ELISA

Cell based phospho ELISA test

Cell based phospho ELISA test

This assay is a direct format to allow detection of phosphorylated EGFRs, JNK, ERK, STATs, phosphotyrosine, and other signaling molecules on adherent cell lines. It is available for human, mouse and rat proteins.


Access to our selection of Cell-based-phospho ELISA Kits here!



Speed ELISA test

Speed ELISA test

#4- Speed ELISA

This sandwich based ELISA has a compressed work flow and only requires 3 hours in total processing time. It is available for human, mouse and rat targets, from cytokines to growth factors and soluble proteins.

Access to our selection of Speed ELISA Kits here!

EIA Format ELISA test

EIA Format ELISA test


#5- EIA Format

This assay is competition based and detects adipokines and peptide hormones for which no antibody pair exist; it is suitable for plasma, serum, culture media, lysates or clarified body fluids from human,


mouse and rat species.


Access to our selection of EIA Kits here!


#6- IQ ELISA FormatIQELISA-Detection


This new ELISA format combines the specificity and ease of use of an ELISA with the high sensitivity of Real Time PCR.

Now available for VEGF, IFN-Gamma, PDGF-BB and HGFR.





Why not outsourcing your ELISA tests to a Raybio’s Certified Service Provider like tebu-bio?

If you’re not familiar with any of these formats, or need to externalise your assay to speed up your research, tebu-bio’s laboratories based near Paris can do the experiment for you. They will offer advice on sample preparation and storage, as well as sensitivity and range of the assay you select.tebu-bio is a Certified Services Provide for RayBiotech ELISA tests and arrays (incl. Quantibody array).

A dedicated project manager will be your direct contact for optimal experimental design and regular reporting in full confidentiality. It’s cheaper than you think! Discover our lab services related to Biomarker validation, Protein profiling and quantification here!

Looking for specific ELISA test?

If this ELISA service is of interest to you, or if you need some advice in choosing the right assay for your project, just ask me your questions below, I’ll be pleased to help. Alternatively, you can use our easy to use ELISA test search engine, and select your target species, application and format.

Popular ELISA tests

Target Cat. Nr
TNF-alpha Human ELISA Kit ELH-TNFa-1
IL-1 beta Human ELISA Kit ELH-IL1b-1
TGF-Beta1 Human Elisa Kit ELH-TGFb1-1
Human OX40 ELISA ELH-OX40-1
Written by Isabelle Nobiron, PhD
Isabelle is a Product Manager at tebu-bio, and also the company's ISO Quality Manager.