What is a SimpleStep ELISA?

• Single-wash, easy protocol reduces assay time to 90 minutes or less
• High sensitivity, specificity, and reproducibility from superior antibodies
• Fully validated in biological samples
• ​96-wells plate breakable into 12 x 8 wells strips
• 10-pack sizes and 384-well formats now available

SimpleStep ELISA^® kits are sandwich ELISA assays that we have developed to be as simple and effective as possible. Unlike traditional sandwich ELISAs, which take more than three hours to run, SimpleStep ELISA kits generate data in just 90 minutes without compromising performance, using only one wash step and recombinant antibodies for exceptional specificity.

How does SimpleStep ELISA work?

• Figure 1. With SimpleStep ELISA kits, an analyte-capture and detector antibody sandwich complex is formed in solution, which binds to the microplate via an affinity tag attached to the capture antibody in the sandwich pair.

A conventional sandwich ELISA uses a pair of antibodies that bind to at least two distinct sites on the target protein. The capture antibody is pre-coated onto the well of a microplate and selectively binds to the target protein. After a wash step, the detector antibody is added and binds to a second site on the target protein to form a sandwich complex.

SimpleStep ELISA kits streamline this process using our CaptSure™ system, a semi-homogeneous system that forms the antibody-analyte sandwich complex in solution in a single step. In just one incubation and wash step, the complete sandwich complex forms in the well and is anchored to the plate with an immunoaffinity tag (figure 1).

Your ELISA upgrade

Competitive SimpleStep ELISA®

Our competitive SimpleStep ELISA^® kits offer you the same benefits in speed and simplicity as our sandwich SimpleStep kits, with the same streamlined protocol and 90-minute run time.

Using our competitive SimpleStep ELISA^® kit, you’ll be able to detect small molecules and hormones not measurable using a sandwich ELISA^® due to the availability of only one epitope or antibody binding site.

Our competitive SimpleStep ELISA® kits use an affinity tag recognized by the monoclonal antibody used to coat our SimpleStep ELISA^® plates. This approach to competitive ELISA^® allows the formulation of the antibody-analyte complex in a single step, saving you significant time at the bench.

In the antigen-down format, the labeled antigen and the unlabeled antigen from your sample compete to bind to the HRP-tagged capture antibody. In the antibody-down format, the HRP-tagged antigen and the unlabeled antigen from your sample compete to bind to the capture antibody. The component labeled with the CaptSure™ affinity tag, either antigen or antibody, binds to the SimpleStep ELISA® plate. A decrease in signal indicates the presence of antigen in the sample when compared to wells containing only the labeled antigen.

Why do we use recombinant antibodies in our SimpleStep ELISA kits?

As ELISAs rely on antibodies to detect the target antigen, the antibodies used must be highly specific.

Recombinant monoclonal antibodies are produced in vitro using high-yield expression vectors, which provides several advantages, including:

• Highest level of consistency and unrivaled reproducibility
• Improved sensitivity and confirmed specificity
• Ease of scalability and continuity of supply

The use of recombinant monoclonal antibodies in our SimpleStep ELISAs, where possible, means you can achieve reliable and reproducible results from the first attempt.

These matched antibody pairs are optimized for use in physiologically relevant sample types, such as plasma and serum. They are also available in carrier-free and conjugation-ready formats, along with individual capture and detector antibodies, to enable flexibility and consistency throughout your workflow.

Uncompromising on sensitivity

SimpleStep ELISA kits drastically reduce assay times without compromising on performance. We compared the sensitivity of SimpleStep ELISA with that of the next best competitor brand of ELISA kits for 75 popular human and mouse proteins. In 56 out of 75 cases, SimpleStep ELISA shows superior sensitivity (figure 2).

Calibration against international standards

All secreted serum or plasma-based targets are tested and fall within World Health Organization blood reference ranges. When available, the SimpleStep ELISA kits are calibrated against known NIBSC international standards and include a conversion factor for data comparison.

Calibrating against an international standard enables comparison to data sets generated from different kits but calibrated against the same standard. Additionally, every SimpleStep ELISA^® kit is validated using multiple biological samples for assay specificity.

Examples of NIBSC conversion factors for our SimpleStep ELISA kits

High-throughput screening (HTS) and SimpleStep ELISA

Our HTS tools and reagents are specially developed to ensure that you get rapid, reproducible, and reliable results with every screening run you carry out. All of our SimpleStep ELISA^® kits can be adapted to a  384-well plate format using the 384-well SimpleStep ELISA^® Microplate. Designed for use with automated liquid handling systems, these pre-coated plates help scale up your research in a flexible solution usable with many standard plate readers.

Immunoassay results 4x faster

With our 384-well SimpleStep ELISA^® kits you can process 4 times more samples than a traditional ELISA format in the same 90 minutes.

We’ve launched a growing range of 90-minute 384-well SimpleStep ELISA^® kits. Our 384-well SimpleStep ELISA^® Microplates and 384-well SimpleStep ELISA^® kits use the same quick and simple one-wash protocol as our standard kits but require less of your sample – just a quarter of the volume required for a 96-well SimpleStep ELISA kit.

We've validated over 280 of our SimpleStep ELISA^® kits in 384-well format. See our most popular 384-well kits below:

Analyte range

We currently have SimpleStep ELISA^® kits to almost 700 targets, including popular proteins like PD-L1, GFP, and fibrinogen, and are continually adding more to the catalog.

SimpleStep ELISA^® kits for the top 10 most popular ELISA targets:

* also available for rabbit, hamster, dog and guinea pigs samples

** also reacts with Rhesus Monkey

*** multispecies