5 Tips to Keep in Mind For Selecting the Most Appropriate ELISA Kits
Researchers use the ELISA kit because of its many advantages. The assay is cost-effective, popular, convenient, simple, and easy to execute. ELISA kits can detect substances in liquid samples using antigens and enzymes, such as tissue lysate, plasma, and culture supernatant.
Considering ELISA’s effectiveness and convenience, it has vast and significant applications in various aspects. For instance, you can use it when conducting biomedical research and disease diagnosis, among many other industries. ELISA can detect any molecule, including nucleic acid, protein, carbohydrates, lipids, and much more.
Most scientists and researchers believe that it is one of the most versatile and popular immunoassays available in the market today. Due to its popularity, you will find that there are various commercial ELISA kits in the market.
But you need to know that not all ELISA kits function the same way. Shifting between masses and selecting an appropriate ELISA kit is a dream come reality to many researchers. Below are a few essential pointers that you should keep in mind when choosing an excellent immune-assay kit.
1. Requirement of Sensitivity
Do you know if the concentration in your target analyte in your sample is high or low? Using a kit with broad range detection is ideal if you are not aware of the level of concentrations in your specimen. However, if you are confident that the concentration level in your sample is low, you require ELISA kits with high sensitivity detection.
Kits with the coating and blocking technology undoubtedly can achieve minimal background, pictogram level sensitivity, and high signal. ELISA kits need to be extremely sensitive to detect low levels concentrations, such as single-digit pictogram levels. Before purchasing an ELISA kit, you need to ensure you know the assay range and its sensitivity levels. Moreover, if the kit is more sensitive, you should be aware that the working range of the kit will be lower.
2. An Inter-Assay Normalization
Another essential aspect that you need to keep in mind for selecting the most appropriate ELISA kits is the inter-assay normalization. First, you need to be aware that ELISA kits in R&D applications lack international normalization. The antibody dependant defines the results that you will obtain.
And this is by reflecting the level of sensitivity of biomarkers in your sample. To have more chances when looking to reproduce the results of the former assay, testing it with the previous kit is ideal. For better results, you need the exact measurements and quality of the samples. Over time you will find that this tip is helpful in your following projects.
The reproducibility of your tests would be more effective if you have an internal control system that you can use on all the plates. You need to know that this internal control is directly related to your assays. Diagnostics assays have a normalized sample preparation, making it hard for research kits to provide internal control.
3. Kit Specificity
Realizing that a strong signal can be a false positive result is annoying and frustrating when using an ELISA kit. It does this when cross-reactivity occurs. Instead of the antibodies attaching themselves to the target analyte, they do so in some other molecule.
Knowing what you want to determine for your sample is an essential aspect of selecting an ELISA kit. Doing this will help you understand what kit is appropriate and which category will work best for your sample. ELISA kits have four general categories, competitive, direct, sandwich, and indirect.
Direct ELISA is the appropriate kit for you if you are looking to get a faster detection of immune response to an antibody. You do not care much about its signal amplification. On the other hand, an indirect ELISA kit is most suitable if you plan for great flexibility and better sensitivity. Doing this helps you to detect the amount of antibody concentration in your sample.
And you might wonder what you will do if you have a complex sample? That should not be an issue, and the sandwich ELISA is the ideal kit for you. It is famous for its specificity, high sensitivity, and flexibility, and you will not need to purify your antigen.
Samples with small antigens or antibodies competitive ELISA should be the go-to go kit. Competitive ELISA can detect very micro antigens that seem difficult for two antibodies to bind together. Assay format is an essential tip that you need to look out for when selecting suitable ELISA kits.
5. The Financial Implication
It is with no doubt that most people consider the cost of an ELISA kit before selecting. Cost is a vital aspect since you are putting in your most significant investment time. Quality is an essential factor to consider, but budget is another important aspect for selecting the most appropriate ELISA kits.
But before purchasing one, you need to have an insight into the number of tests one kit can provide. In addition, even the amount of sample the kit will require in conducting an assay. You need to consider ELISA kits with removable strips, and this helps you use one kit at a time and store the rest for later.
Moreover, if you are a value-conscious customer and have experience using ELISA kits, you can use an antibody pair. Most researchers will confirm that this is a low-cost ELISA solution since it comes with a DIY ELISA matching antibody pair. Also, this particular kit has the necessary core reagents that develop and run the ELISA test.
Summary
Most people are looking for appropriate ELISA kits for use. But they are not sure about the top things they need to consider before selecting a good kit. Kit sensitivity, assay format, inter-assay normalization, kit specificity, and budget are a few things you need to keep in mind when selecting a kit for your sample. Doing this will guide you into defining your project plan; you will build a personal multiplex assay and choose the best ELISA for your model.
