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Highly sensitive 90-minute ELISAs, with just one wash step and no compromise on performance.
Contents
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 by using a semi-homogeneous format wherein the antibody-analyte sandwich complex is formed 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. |
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 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 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.
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
Product | AbID | Standard ID | NIBSC conversion factor |
---|---|---|---|
Human Erythropoietin (EPO) | ab274397 | 11/170 | 0.001 |
Human alpha 1 Antitrypsin (SERPINA1) | ab189579 | 05/162 | 0.3649 |
Human BMP2 | ab277085 | 93/574 | 0.00024 |
Human C-Peptide | ab260064 | 13/146 | 1.077 |
Human Flt-3 ligand | ab254507 | 96/532 | 0.0014Tabl |
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. Our SimpleStep ELISA kits are adaptable in a 384-well plate format and can be used with automated liquid handling systems. These plates help scale up your research and provide a flexible solution that can be used on many standard plate readers. The 384-well plate SimpleStep ELISA uses the same quick and simple one wash protocol as our standard SimpleStep ELISA kits but requires less of your sample to run each plate.
Find more technical information about our 384-well SimpleStep ELISA Microplate here
Find out more about our ELISA-based high-throughput screening solutions here
SimpleStep ELISA kits and the SPECTROstar®Nano
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.
See how:
Application note 2: Immuno-oncology biomarker measurement with high sensitivity and speed using SimpleStep ELISA® kits and SPECTROstar® Nano.
Discover why:
Application note 3: Fast and accurate detection of Alzheimer’s Disease targets with SimpleStep ELISA® kits and SPECTROstar® Nano
Learn more about:
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:
Target | Human | Mouse | Rat |
IgG* | ab195215 | ab189578 | |
Albumin | ab207620 | ab235642 | |
TNF alpha | ab181421 | ab208348 | |
IL-6 | ab178013 | ab222503 | ab234570 |
IFN Gamma | ab174443 | ||
IL-1 beta | ab214025 | ab197742 | |
GFP | ab171581*** | ||
Alpha 1 antitrypsin | ab189579 | ||
Pro collagen I alpha | ab210966 | ab210579 | |
Fibrinogen | ab241383 | ab213478 |
* also available for rabbit, hamster, dog and guinea pigs samples
** also reacts with Rhesus Monkey
*** multispecies
Find your SimpleStep ELISA® kit
ELISA guide
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