The enzyme-linked immunosorbent assay ELISA is a highly sensitive method to quantify the presence of a ligand (usually a protein). Enzyme-linked antibodies are used to bind the analyte, after which the addition of a substrate will trigger a chemical reaction that will convert a colorless reporter molecule to a colored product for detection. This widely applied assay offers high specificity and high sensitivity for a wide variety of analytes in different kinds of samples. Since it was invented four decades ago, ELISA has been applied to many fields such as biomedicine, environmental studies, and food quality. During the past 30 years, Bertin Bioreagent has been developing a strong ELISA assay expertise, based on the use of the Acetylcholinesterase enzyme (AChE) technology. We have designed and validated more than 100 ELISA assays for biomarkers in fields such as metabolic diseases, pain, or inflammation.
We invite you to read the following text to discover helpful tips on how to choose the best sample preparation methods for ELISA assays.
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Tips and tricks
Sample preparation and collection are among the most crucial steps in the ELISA workflow to obtain accurate and reproducible results. The goals of these steps include stabilizing the molecules of interest, and also extracting and resuspending them in an optimized matrix to allow for the detection of the target molecule.
During sample collection, it is essential to maintain the integrity of the sample until the ELISA assay can be performed. Bertin Bioreagent offers a wide variety of innovative collection tools to ensure the stabilization of your sample.
Our Sampling tubes with Indomethacin can be used to collect blood while preventing the ex-vivo formation of Thromboxane B2 formation and other prostaglandins, thanks to the cyclooxygenase (COX) inhibitor Indomethacin.
Our Sampling tubes with Butylated Hydroxytoluene (BHT) have been shown to inhibit lipid peroxidation and the formation of other ROS species.
Animal tissues and cells samples have to be converted to a liquid form to be compatible with ELISA assays. Bertin Technologies has designed the Precellys, a range of homogenizers powered by 3D bead-beating technology, to offer researchers a versatile and high-throughput solution for their sample preparation needs. The Precellys lysing kits can be used to homogenize virtually any kind of sample efficiently and uniformly.
Additionally, our Protein Safe Precellys lysing kits contain a cocktail of protease inhibitors to protect your samples against degradation throughout the whole homogenization process.
Following the collection, many factors can affect the concentration of the targeted analyte. Bertin Bioreagent has developed several inhibitors cocktails to stabilize the concentration of relevant biomarkers in the sample to physiological values.
Our Angiotensin II Inhibitor Cocktail has been designed by Bioreagent to stop sample degradation. This cocktail offers a convenient and reproducible solution to stabilize Angiotensin II concentration in blood samples for long-term storage.
Insulin measurement in serum samples presents many challenges related to hemolysis, circulating anti-insulin autoantibodies, and the reactivity (or lack thereof) of rapid or long-acting pharmacological insulin analogs in serum. Hemolyzed samples contain an insulin-degrading enzyme, and should only be analyzed (1) if they can be handled at 4°C within 2-3h after collection or (2) an insulinase inhibitor cocktail has been added in the blood collection tube to prevent insulin degradation. Bertin Bioreagent has developed the Insulinase Inhibitor Cocktail to offer researchers a turnkey solution for insulin measurement in serum samples. Our Insulinase Inhibitor Cocktail has been validated with the Bertin Bioreagen Insulin (mouse, rat) ELISA kit # A05105.
Matrix preparation products
Our CGRP affinity sorbent contains mouse monoclonal anti-CGRP antibodies immobilized on a solid matrix. This product has been developed as a convenient tool to prepare CGRP-free plasma, which can be used as a sample matrix blank (SMB) to dilute standards and samples prior to ELISA analysis.
Appropriate sample preparation for CGRP ELISA kits is key for the accurate quantification of the peptide. The following diagram illustrates the options available according to the type of matrix. For a detailed protocol, please read the user’s manual of the kit.