ELISA Plate Readers

What is an ELISA microplate reader?

A microplate reader is an instrument used by researchers to detect and analyse a range of biochemical changes. An ELISA plate reader can read and analyze multiple wells at once at a range of wavelengths, the ELISA plate reader allows researchers to improve efficiency and save operational time and costs while automating a data analysis for a range of applications.

How does an ELISA plate reader work?

An ELISA plate reader works by detecting light at a specific wavelength. For the final steps in the ELISA assay, TMB (3,3',5,5'-Tetramethylbenzidine) is converted to a blue coloured solution by HRP and finally converted to a Yellow Coloured solution following the addition of stop solution. The amount of colour in the well is proportional to the amount of analyte that is in a sample.

Once the plate is placed in the ELISA plate reader the absorbance of each sample is read at 450nm to determine the amount of analyte in each well. Researchers can then plot the value of their samples vs known values of their standard curve to calculate the amount of analyte they have in each sample.

96 well ELISA plate readers

When carrying out an ELISA assay you will perform your assay in a 96 well plate to measure the amount of analyte in each sample. The formatting of your plate will consist of an 8 point standard curve and blanks in duplicate. This will take up 20 wells on your 96 well ELISA plate. This the researcher to measure 38 samples in duplicate on their 96 well ELISA plate. 

There are a range of ELISA plate readers on the market that will allow you to measure your ELISA assay including SPECTROstar Nano from BMG LABTECH, plus any plate reader from Bio-Rad, Thermo, Tecan, Molecular Devices or Biotek. Each platform comes with it’s own dedicated plate readers software to allow researchers interpret their data.

Company Platereader




PowerWave HT Microplate Spectrophotometer




xMark™ Microplate Absorbance Spectrophotometer

Plate Reader Settings to measure your ELISA assay

Depending on the ELISA plate reader that you have, settings may vary. However, the following settings may apply when reading your sample:


Optic settings

Absorbance spectrum, Endpoint test

Wavelength 400-700nm

General setting

Number of flashes: 45

Setting time: 0.2s

ELISA plate layout map

Recording positions in your ELISA plate

When carrying out an ELISA assay it is important to record the positions of the standards, samples, controls and blanks that will be added to the plate and for further analysis by the ELISA plate reader. Above is a standard layout that most researchers use when running their standards and blanks in duplicate. This allows researchers to analyse 40 samples on a 96 well ELISA plate.