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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.
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. See the video of how SimpleStep ELISA works.
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).
|SimpleStep ELISA® offers
|What it means for you
|High sensitivity, reproducibility, and specificity from recombinant antibodies
|Consistent and reliable results from the first attempt
|Validation on biological samples
|Guaranteed reaction with the endogenous protein
|Single-wash, quick (90 minutes) standardized protocol
|Less risk of error with the same fast and easy protocol across all SimpleStep ELISA® kits
|96-wells plate breakable into 12 x 8 wells strips
|No waste. Possibility to use the same kit for several smaller experiments
|10-pack sizes now available
|Guarantees same lot
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 antigen labeled with a CaptSure™ affinity tag 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 unlabeled antigen from your sample compete to bind to the capture antibody labeled with a CaptSure™ affinity tag. The CaptSure™-labeled antigen or antibody binds to the SimpleStep ELISA® plate. A decrease in signal indicates the presence of the antigen in samples when compared to assay wells with labeled antigen alone.
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:
The use of recombinant monoclonal antibodies in our SimpleStep ELISA®s, 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 the individual capture and detector antibodies, to enable flexibility and consistency throughout your workflow.
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).
Figure 2. We compared SimpleStep ELISA® kit sensitivity with a competitor assay across 75 popular protein targets and calculated the ratios. SimpleStep ELISA® was more sensitive on average than a competitor ELISA kit.
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
|NIBSC conversion factor
|Human Erythropoietin (EPO)
|Human alpha 1 Antitrypsin (SERPINA1)
|Human Flt-3 ligand
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.
We’ve also 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.
To get the best results from your SimpleStep ELISA® kits we recommend that you run your plates using the SPECTROstar®Nano. Reading microplates with the SPECTROstar®Nano provides outstanding data quality while offering the convenience of linking data analysis to MARS data analysis templates for immediate and accurate results.
Discover how you can use the SPECTROstar®Nano in combination with SimpleStep ELISA® kits in this package of three application notes.
Application note 1: Highly sensitive ELISAs in 90-minutes: SimpleStep ELISA® kits and the SPECTROstar® Nano.
Application note 2: Immuno-oncology biomarker measurement with high sensitivity and speed using SimpleStep ELISA® kits and SPECTROstar®Nano.
Application note 3: Fast and accurate detection of Alzheimer’s Disease targets with SimpleStep ELISA® kits and SPECTROstar®Nano
Learn more about:
SimpleStep ELISA® kits for the top 10 most popular ELISA targets:
|Alpha 1 antitrypsin
|Pro collagen I alpha
* also available for rabbit, hamster, dog and guinea pigs samples
** also reacts with Rhesus Monkey