ELISA and Western blot are two of the most common molecular techniques used to detect and quantify analytes. Both these techniques have similar principles of antibody-antigen binding, but they have different advantages and applications. The current article explores the difference between ELISA protein assay and western blot technique and is a guide for immunoassay services to choose the right method for the research. Whether it’s ELISA services or PCR services, robust ELISA method development and validation is critical for reliable, reproducible, and accurate results.
Difference between ELISA immunoassay and western blotting technique
ELISA services perform ELISA in microplates in a simple and highly sensitive format to measure the presence of specific analytes in study samples. ELISA immunoassays are often quantitative and qualitative assessments of proteins in different matrices such as cell supernatants, serum, plasma, and urine.
Western blotting can characterize and detect specific proteins in study samples. The basic technique involves protein separation using electrophoresis followed by transferring it onto a membrane and using antibodies specific to the analyte of interest. If the target analyte is present in the sample, specific antibodies will bind it. Finally, the secondary antibody binds to the primary antibody and detects it through fluorescence or chemiluminescence methods.
Understanding the advantages and disadvantages of both these methods, ELISA assay development services can make informed decisions to choose an ideal method for their experiment. Both techniques can quantify and detect proteins in study samples. However, each has its unique characteristics and limitations. Let us understand each of these techniques in more detail.
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ELISA protein assay
ELISA immunoassays can detect very small quantities of target analytes or low or rare abundance proteins. It can measure the exact quantity of target analytes in study samples, proving a valuable tool to monitor changes or determine analyte concentrations. ELISA assays are rapid and easy to perform with minimal sample preparation. Hence, it is an ideal choice for routine immunoassays.
However, ELISA assays pose some limitations. Traditional ELISA assays are single-plex and can detect only one analyte at a time. Hence, scientists find it challenging to measure multiple analytes simultaneously in a single assay volume.
Western blot technique
Western blot analysis is highly specific in resolving proteins based on their molecular weight. Hence, it is ideal for detecting protein modifications and differences between experimental setups. Western blot technique is valuable for analyzing complex mixtures and analytes over a wide range of different applications. This technique can identify low levels of analytes present in complex mixtures. Besides, a significant advantage of the Western blot technique is that it can detect proteins based on their size and can help reveal impurities or protein modifications.
However, the Western blot technique is a multi-step method and can take multiple hours to overnight for one essay. Besides, it is less sensitive than ELISA assays.
Hence, while choosing between western blot and ELISA immunoassay, one must consider individual experimental needs and the intended application of a protein assay. Both techniques are vital bioanalytical methods, and hence, selecting one will depend on specific requirements.
