Congratulations, your proof of concept has worked. You've validated your assay design, optimized your protocol and troubleshot the issues. Now, you’re ready to scale up to high-throughput screening.

That’s when the headaches can start scaling up, too.

“It all started off perfectly,” shared one researcher about a transition during drug development. “The problems began when we tried to verify reproducibility.”

Scaling up to high-throughput screening can be time-consuming and labor-intensive, and often demands painful trade-offs between scale, sensitivity, specificity and simplicity. One of the biggest challenges of a transition is ensuring that results stay reproducible from lab to lab.

But what if you knew from the start that you could easily scale from low to high throughput and from lab to lab with the same simple protocol, the same high-quality results – and no trade-offs?

Introducing the fully validated 384-well SimpleStep ELISA® kit

Abcam’s 96-well SimpleStep ELISA® kit revolutionized lab workflows by being the first to generate data in just 90 minutes or less, using a simple, single-wash protocol. A traditional ELISA takes around four hours. Since 2012, our kits have saved researchers over 607,500 hours at the bench – that’s over 69 years.

SimpleStep uses our innovative CaptSure™ technology, 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.

As well as speed, SimpleStep ELISA® kits deliver exceptional sensitivity, specificity, flexibility and reproducibility, thanks to our robustly validated recombinant antibodies.

All these advantages are now available in a 384-well SimpleStep ELISA kit on a standard plate that's compatible with automated liquid handling systems and requires just a quarter of the sample. Together, our 96-well and 384-well kits enable easy, seamless scaling.

“You can develop your assay using a 96-well kit, move directly into the 384-well plate... and just crank through your data samples.”

Jeff Monette, Abcam’s Head of Immunoassay Platform Innovation

Seamless scaling with consistent, comparable results

Our 384-well SimpleStep ELISA® kits are fully validated and seamlessly interchangeable with their 96-well counterparts. They use exactly the same precision recombinant antibody pairs, guaranteeing you the same accuracy, precision, linearity, and detection limits.

The same simple, rapid protocol
The 384-well SimpleStep ELISA® uses the same 90-minute mix-wash-read protocol as the 96-well kit, using the same buffers and materials. This means no adaptation or revalidation of your assay is needed during scaling, allowing you to make a simple handover to a new team or lab, and saving you time while running samples.

Lab-to-lab reproducibility is assured
The 384-well SimpleStep ELISA® kit is fully validated to guarantee low intra-assay variability, so you can be confident in achieving comparable results across teams and labs, no matter where or when they’re generated.

Get more done with less sample
The 384-well SimpleStep ELISA® kit uses one quarter of the sample volume of the 96-well kits, so you can achieve more with less. Using just 12.5 microliters instead of 50 is ideal for measuring multiple targets with limited sample volume, using precious samples more efficiently, and lowering costs.

“The SimpleStep ELISA® kit cuts my work day by many, many hours, and I’m able to get the results to my team in a really quick and convenient way.”

Mary Wozniak, Senior Research Associate, Arkuda

“One of the fantastic things about our 384-well SimpleStep ELISA® kits is that we validate them against our current 96-well kits, and they're absolutely interchangeable,” says Jeff Monette, Abcam’s Head of Immunoassay Platform Innovation.

“So you can develop your assay using a 96-well kit, which is extremely friendly for manual operation. And when you're happy with the results and optimized protocol, you can move directly into the 384-well plate, place it on your liquid handler and your automation, and just crank through your data samples.”

Supporting pre-clinical and clinical research
Our 384-well Human Albumin and Human ALT SimpleStep ELISA® kits allow assessing liver toxicity on a larger scale using less sample volume.

Thanks to their one-wash protocol, SimpleStep ELISA® kits can also be used as orthogonal screening methods to confirm hits identified by homogeneous immunoassays, eliminating false positive results.

Our spectrum of immunoassay solutions gives you total flexibility
The core components that power our SimpleStep ELISA® kits are available individually in multiple formats to give you extra flexibility, including carrier-free antibody pairs, individual carrier-free capture and detector antibodies, conjugation kits and protein standards. Our 1,700+ carrier-free antibody pairs – the world’s largest range – are conjugation-ready and adaptable for your own unique assay design, with 330+ protein standards supporting validation.

Our 96-well SimpleStep ELISA® kits also come in packs of 10 that are easier to integrate into your automation, and easier to store, with bulk bottles of reagents and reduced packaging.

All powered by recombinant technology
Our recombinant antibodies deliver the highest specificity, sensitivity, and consistency, overcoming traditional problems such as low reproducibility and genetic drift by cloning specific antibody genes into vectors in vitro.

We also use biophysical characterization to confirm antibody identity at a molecular level, winning us a Highly Commended Digital Science Tool of the Year award from CiteAb in 2024. This guarantees lot-to-lot consistency, so you can be confident of supplies and consistent results again and again over long-term projects. We now offer 32,000+ recombinant antibodies in the world’s largest range, so you can find exactly what you need.

Our scientific experts are on your team
Our global, multilingual Scientific Support team is made up of scientists with real-world lab experience who stay on the cutting edge of research through ongoing training. They’re ready to help and advise you on the latest protocols, techniques, and products, and can help you troubleshoot and resolve challenges in designing and running your experiments.

Immunoassays that keep you moving forward
At Abcam, we’re known for our exceptional antibodies. We use that high quality to deliver flexible immunoassay solutions you can scale and adapt without sacrificing sensitivity, specificity, or simplicity – and keep your research moving forward.

CaptSure is a trademark of TGR BioSciences Pty Ltd, Australia

====

The fallibility of antibodies is not news; there are widespread reports of researchers failing to replicate or having to retract findings because of poorly characterized antibodies or differences between batches1.

These headline cases are symptomatic of a much more pervasive and pernicious problem: researchers are losing confidence in antibodies.

It’s a familiar headache for Will Howat, vice-president of validation and technical quality at Abcam, a global producer of research reagents. As a former molecular pathology team leader at a pharmaceutical company, he recalls having to modify assays every time he got a new antibody batch. “I’d have an assay set up, and have titrated the antibody in my system across different tissue types, only to find it would no longer work the same with the next batch of antibody. So, research stops while you titrate again. It’s a lot of work.”

What adds to this problem is a lackadaisical approach to antibody reporting across the scientific literature, something Bandrowski is working hard to change.

“In order to define good antibodies, or even bad antibodies, one needs to first identify them,” says Bandrowski. “The unfortunate problem is that authors do not identify the antibodies they use an estimated 20% to 50% of the time.”

Initiatives such as the Antibody Registry² and the Research Resource Identification Initiative³ have already markedly improved antibody identifiability. However, if those antibodies are not properly characterized, results may still be questionable.

Taking an antibody's fingerprint

Using recombinant antibodies, which are produced from a specific genetic sequence that does not vary over time, removes the variability inherent with hybridoma-produced antibodies, but quality control (QC) is still essential to ensure no natural product variation creeps in.

Antibodies produced for therapeutic purposes are subjected to rigorous biophysical QC methods, including liquid chromatography-mass spectrometry, dynamic light scattering and high-performance liquid chromatography, to ensure they meet regulators’ good manufacturing practices for biopharmaceuticals. But there are no agreed QC standards for research-use only (RUO) antibodies.

“It’s critical to know that you’ve got the confidence of a pure antibody product and a reproducible supply”

Will Howat, Vice-President of Validation and Technical Quality, Abcam

The IWGAV recommended application testing and knock-out validation as key pillars for the validation of RUO antibodies4, which Abcam has been doing for many years. The company has now raised the bar even higher by introducing stringent biophysical QC standards across its catalogue of recombinant rabbit monoclonal RUO antibodies, with 75% tested to date.

“Biophysical characterization is not about how the antibody works in an assay, it’s about its identity,” says Howat. “By confirming its mass, its aggregation, its purity — all of which are unique to a specific antibody — we can pull those features together into an antibody fingerprint, so you know the reagent is exactly what it should be.”

Abcam is taking on these additional tests in order to help overcome the reproducibility crisis. “Whether you’re a pharmaceutical scientist who needs to seamlessly move from discovery to development, or a facility manager concerned by consistency of product quality and performance, it’s critical to know that you’ve got the confidence of a pure antibody product and a reproducible supply,” says Howat.

For all antibody research, starting with identified, quality antibodies is paramount, according to Bandrowski, and should always be followed by confirmation that the antibody performs as intended. “If you’re not validating your reagents, you can’t be confident in what they’re telling you,” she warns. “And if you publish those data, you can’t be confident in what you’re contributing to the scientific record."

References

1. Baker, M. Nature 521, 274‒276 (2015).
2. Bandrowski, A. et al. Nucleic Acids Res. 51, D358‒D367 (2023).
3. Bandrowski, A. et al. J. Comp. Neurol. 524, 8‒22 (2016).
4. Uhlen, M. et al. Nature Methods 13, 823‒827 (2016).

You may also be interested in…