Commercial ELISA Kits are being widely viewed as the perfect solution for biomedical analysis and various industry research projects. Their convenience, speedy delivery, and versatility make them the most potent immunoassays available in the market today. These enzyme-linked immunosorbent assay (ELISA) kits are also available in every standard, depending on how you want to use them. Given these differences and their varied applications, it might seem challenging to decide which one to choose for your respective lab project. Here, we provide you with the proper steps that can help you select a perfect ELISA Kit suitable for all your intended purposes.
1. Establish the Analytes You Want to be Detected
Some of the commercially available Elisa Kits are designed to measure serum and culture supernatants. This means you have to carefully read through each kit’s use to ascertain if it’ll perfectly suit your lab project.
For instance, Sandwich ELISA kits will serve you best to detect bulky proteins with various epitopes, such as a cytokine. On the other hand, competitive ELISA kits may be just what you’re looking for if your lab project is all about detecting small molecules, such as hapten. Other factors such as the collection method of plasma samples also affect the type of ELISA kit you choose.
When choosing your kit, you also need to be wary of lipid facets and hemolysis in the sample. Taking all these into consideration before selecting your ELISA kit is essential in achieving the results you want. A pre-test trial can also serve as a perfect way to verify the kit’s suitability, compatibility, and sensitivity to your samples.
2. Your Sample Species and Type
The sample species you want to test will determine the type of kit to use. This is why all ELISA kits are validated for species that show homology with different sample species. Most of the readily available commercial ELISA kits are commonly used on humans, mice, and rat species.
However, if your lab samples are drawn from unique species like monkeys, you’ll have limited ELISA kits to choose from. This will require you to go for any available kit that has been validated for a species showing similar homology with your sample species. Homological sequences with more than 90 percent identity often indicate higher chances of cross-reactivity.
3. The System of Detection You Want to Use
Enzyme-linked immunosorbent assay kits come with various detection systems. The kits employ the technique of obstructing the analyte on the surface, marking the enzyme, and matching the substrate. The detection systems widely used today include luminescent, colorimetric, and fluorescent techniques.
Therefore, it’s essential first to select a suitable enzyme and a matching substrate. In addition, your kit choice should reflect on the enzyme-substrate for cross-reactivity of the substrates. This way, you’ll have a kit that serves its functions correctly, without any challenges.
4. The Kits Recovery and Linearity Data
Conducting recovery and linearity experiments serves as a primary way to assess and monitor the performance of ELISA kits. If you’re lucky enough, you’ll find this data well outlined in the kit’s specifications. The information also comes along with other vital factors such as sensitivity and the kit’s dynamic range.
Recovery data will help you understand if the analyte detection is affected by any variances in sample conditions. Usually, an Elisa Kit with a higher recovery is considered an ideal choice for complex experiments.
On the other hand, the extent of dilution’s linearity defines the dose-response of the analyte in any given diluent. The condition is termed perfect if the concentration of the samples is found similar for all dilutions. ELISA kits may have different parameter data depending on the manufacturer. The best way to identify one that will suit your lab projects is to compare the data from the experiments carefully.
5. Sensitivity and Specificity Requirements
ELISA Kits are designed for a wide range of detection capabilities. For instance, those offering a more comprehensive detection range will suffice if you don’t know about your sample’s analyte concentration. In addition, ELISA Kits offering higher sensitivity come in handy when your sample has a low analyte concentration. However, if your samples have high analyte concentrations, you can dilute them to fit in the kit’s adaptation range.
Other challenges that come with the choice of ELISA kits are cross-reactivity and obtaining a false-positive result. This is a common tendency when the antibodies attach to the wrong molecule instead of the intended analyte. Such interferences are expected if the sample components alter the antibody-antigen interaction. The outcome is often obstruction of the antibodies from finding the right analyte. That’s why you should always check on the cross-reactivity data from the manufacturer before selecting any ELISA kit for your lab projects.
Bottom Line
The enzyme-linked immunosorbent assay kits are recognized worldwide for their benefits in biomedical experiments. Their reliability and high accuracy levels mean you can always rely on them for complex lab projects and other research studies. But before buying one, explore the above-shared tips to get an insight into choosing one that will serve the proper purpose for your lab project.
