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Benefit from our expertise to save time and resources on your projects. Based in Europe, our laboratory combines proximity and efficiency to successfully drive your research with you and for you.
Life Sciences research is demanding, leaving no room for inefficiency.
That's why we've developed tailored solutions that go beyond the typical distributor role, allowing you to focus on what matters most: achieving meaningful results and driving your research to success.
Discover how our procurement solutions can support you:
Accelerate Your Research with Tebubio's Contract Research Services: Our complementary platforms are designed to deliver seamless solutions, guided by a dedicated PhD-level project manager as your single point of contact.
Benefit from our expertise to save time and resources on your projects. Based in Europe, our laboratory combines proximity and efficiency to successfully drive your research with you and for you.
Life Sciences research is demanding, leaving no room for inefficiency.
That's why we've developed tailored solutions that go beyond the typical distributor role, allowing you to focus on what matters most: achieving meaningful results and driving your research to success.
Discover how our procurement solutions can support you:
I know, this might sound like the most obvious advice ever. But trust me, it’s worth emphasizing! Take a moment to double-check the protocol that comes with our kits. Why? Because our protocols aren’t one-size-fits-all. They differ not just between applications (like ELISA, ELISpot, and FluoroSpot) but might be specific to the analyte. For instance, the recommended concentrations of capture and coating antibodies can vary between kits.
Sometimes, we tweak a protocol to accommodate batch-specific updates (like the ELISA standards) or introduce improvements, such as our updated sample dilution recommendation for our Human ApoB ELISA.
By following the provided protocol, you’ll reduce the risk of troubleshooting. Speaking of which, check out our ELISA troubleshooting guide for extra tips. Most importantly, sticking to the protocol ensures reproducibility – central to reliable, consistent results across experiments.
Looking for a more in-depth breakdown of our ELISA protocol? Explore our ELISA step-by-step guide for all the details!
Your choice of plate can make or break your ELISA results, so don’t grab just any plate off the shelf! ELISA plates are specially designed to bind high amounts of protein.
Here’s what to look for:
And one more thing – cell culture plates are a no-go! They’re great for growing cells, but not for ELISA experiments. Stick with plates made for the job, and you’ll set yourself up for success.
If you prefer to skip plate coating altogether, have a look at our ELISA Pro kits which include pre-coated plates.
Before diving into your ELISA experiment, take a moment to ensure you have all the necessary reagents and consumables on hand. Running out mid-assay? Not fun.
Pay extra attention to your enzyme conjugate and substrate: they need to be compatible with your ELISA setup. If you’re using our ELISA Flex kits, remember that substrates are sold separately. But if you’re working with our ELISA Pro or ELISA PathRF kits, good news, these reagents are already included.
Your substrate choice matters. It directly impacts assay sensitivity, so do a quick quality check:
Need help choosing between an ALP- or HRP-based ELISA? Click here for guidance.
The antibody pair in your assay isn’t just another reagent – it’s the key player determining your ELISA’s sensitivity, specificity, and dynamic range. Choose wisely!
Goto TSKT to know more.
At Mabtech, we validate our antibody pairs for each application, ensuring they bind to the native protein (and yes, that’s important – find out why here).
Another crucial step? Finding the optimal antibody concentrations – this can vary depending on your assay or setup. But if you’re using a Mabtech ELISA kit, good news, we’ve already done the titration work for you. Just follow the recommended concentrations in your ELISA protocol, and you’re good to go.
Heterophilic antibodies can disrupt your ELISA by crosslinking capture and detection antibodies, ultimately leading to false-positive results. The outcome? Inaccurate analyte quantification – nobody wants that.
What are heterophilic antibodies?
Heterophilic antibodies bind to other antibodies. In human samples, these can be:
How do heterophilic antibodies affect ELISA?
Heterophilic interference is commonly found in plasma and serum samples across species. Prevalence estimates for HAIA in the general population vary widely, between 1% to 80% (Kricka ). Particularly high concentrations of heterophilic antibodies and rheumatoid factor (RF) are prevalent in samples from patients with conditions such as rheumatoid arthritis, cancer, or chronic infections.
How to reduce interference?
We recommend you trying our ELISA diluent when analyzing plasma and serum samples. Our ELISA PathRF kits have specifically been developed to eliminate heterophilic interference. These kits feature a recombinant backbone in the detection antibody and a specialized RF-block diluent. In validation experiments with RF-containing plasma, ELISA PathRF kits eliminated false-positive results. Click to learn more.
Once your ELISA plate has been read, it’s time to analyze the data. Luckily, most ELISA readers simplify this process, but understanding the basics will help you make the most of your results.
Get to know your reader software
Before diving into the data, take a moment to explore your ELISA reader’s capabilities. Many software programs allow you to create templates, set up automatic calculations, and adjust analysis settings. These features can save you time in future experiments.
Processing OD values
The optical density (OD) values need a bit of cleanup before they’re useful:
Check the raw data
Before trusting your results, take a moment to review the raw OD values. Look for inconsistencies like unusually high or low readings (which may indicate pipetting errors), discrepancies between duplicates (CV values >10%), edge effects, or contamination. If something looks off, consider repeating the assay or excluding problematic data points.
Choose a good standard curve model
The accuracy of your sample concentration calculations depends on the curve fit. Avoid using a simple linear model unless your data truly follows a straight line. Instead, choose a 4-parameter logistic (4PL) or 5-parameter logistic (5PL) regression. These provide better curve fits for ELISA’s typical sigmoidal response curves, and finally more reliable results.
The perfect ELISA setup depends on several factors – your experimental goal, antibody/antigen availability, and analyte size, to name a few. But when in doubt, we always reach for a sandwich ELISA first!
Why sandwich ELISA?
If you're working with low-abundance targets in complex samples (loaded with other proteins), a sandwich ELISA is your best bet for high sensitivity. In contrast, direct and indirect ELISAs may lack the sensitivity needed – or fail entirely – under these conditions.
Why two-step detection?
If sensitivity is key, go for an ELISA with two-step detection – it enhances the signal for more precise measurements. (That said, if you're measuring an abundant analyte like IgG, a one-step detection kit might be the better fit.)
Looking for antigen-specific immunoglobulin detection? Check out this blog post for the best setup options!
If you are looking for more details, kindly visit ELISA Kits.