Sample Preparation and Handling
Expert-defined terms from the Masterclass Certificate in ELISA Assays course at London School of Business and Administration. Free to read, free to share, paired with a professional course.
Absorbance refers to the amount of light absorbed by a sample, which is a critic… #
Related terms include transmittance and optical density, which are used to describe the amount of light that passes through a sample and the ratio of absorbed to transmitted light, respectively. In ELISA assays, absorbance is measured using a spectrophotometer, which is an instrument that measures the interaction between light and the sample. The absorbance of a sample is directly proportional to the concentration of the analyte, allowing researchers to quantify the amount of analyte present.
Accuracy refers to the degree of closeness of a measurement to the true v… #
Related terms include precision and reliability, which describe the consistency of measurements and the ability of a method to produce consistent results, respectively. In ELISA assays, accuracy is critical, as small errors can lead to incorrect conclusions. To ensure accuracy, researchers use calibration curves and controls to validate the results.
Addition of reagents is a critical step in ELISA assays, as it inv… #
Related terms include dilution and concentration, which describe the process of adjusting the concentration of the sample or reagents. In ELISA assays, the addition of reagents must be done carefully, as the ratio of reagents to sample can affect the results.
Analyte refers to the substance being measured in an ELISA assay,… #
Related terms include antigen and ligand, which describe the substance that binds to the analyte. In ELISA assays, the analyte is the target of the assay, and its concentration is measured using a detection system.
Antibody refers to a protein produced by the immune system in response to… #
Related terms include immunoglobulin and immunoassay, which describe the type of protein and the type of assay that uses antibodies, respectively. In ELISA assays, antibodies are used to capture and detect the analyte, and their specificity and sensitivity are critical to the success of the assay.
Antigen refers to a substance that triggers the production of antibodi… #
Related terms include hapten and epitope, which describe a small molecule that can trigger an immune response and the specific region on an antigen that is recognized by an antibody, respectively. In ELISA assays, antigens are used to capture the analyte, and their concentration and purity can affect the results.
Assay refers to a procedure used to measure the concentration or <… #
Related terms include test and measurement, which describe the process of evaluating a sample and the process of obtaining a numerical value, respectively. In ELISA assays, the assay is designed to detect the analyte, and its sensitivity and specificity are critical to the success of the assay.
Automation refers to the use of instruments and software to perfor… #
Related terms include robotics and high-throughput screening, which describe the use of automated systems to perform multiple assays simultaneously. In ELISA assays, automation can be used to perform tasks such as sample preparation and detection.
Calibration refers to the process of adjusting the response of a d… #
Related terms include standardization and validation, which describe the process of establishing a common reference point and the process of evaluating the performance of an assay, respectively. In ELISA assays, calibration is done using calibration curves and controls.
Capture antibody refers to an antibody that is used to bind … #
Related terms include coating antibody and immobilized antibody, which describe the process of attaching the antibody to a solid surface. In ELISA assays, the capture antibody is used to capture the analyte, and its specificity and sensitivity are critical to the success of the assay.
Certification refers to the process of verifying the performance a… #
Related terms include validation and qualification, which describe the process of evaluating the performance of an assay and the process of verifying the performance of an instrument, respectively. In ELISA assays, certification is done by regulatory agencies and manufacturers.
Chemiluminescence refers to the emission of light by a chemical re… #
Related terms include bioluminescence and fluorescence, which describe the emission of light by living organisms and the absorption and emission of light by a molecule, respectively. In ELISA assays, chemiluminescence is used to detect the analyte, and its sensitivity and specificity are critical to the success of the assay.
Coating refers to the process of attaching a capture antibody or <… #
Related terms include immobilization and adsorption, which describe the process of attaching a molecule to a surface and the process of attracting and holding molecules to a surface, respectively. In ELISA assays, coating is done to capture the analyte, and the concentration and purity of the coating reagent can affect the results.
Colorimetry refers to the measurement of the absorbance of ligh… #
Related terms include spectrophotometry and photometry, which describe the measurement of the interaction between light and a sample and the measurement of the amount of light absorbed by a sample, respectively. In ELISA assays, colorimetry is used to detect the analyte, and its sensitivity and specificity are critical to the success of the assay.
Control refers to a sample with a known concentration of analyte,… #
Related terms include calibrator and standard, which describe a sample with a known concentration of analyte and a sample with a certified concentration of analyte, respectively. In ELISA assays, controls are used to ensure accuracy and reliability, and their concentration and purity can affect the results.
Detection refers to the process of identifying the presence or … #
Related terms include measurement and quantitation, which describe the process of obtaining a numerical value and the process of determining the amount of a substance, respectively. In ELISA assays, detection is done using a detection system, which can be based on colorimetry, chemiluminescence, or fluorescence.
Dilution refers to the process of adjusting the concentration of a… #
Related terms include concentration and serial dilution, which describe the process of adjusting the concentration of a sample and the process of making multiple dilutions of a sample, respectively. In ELISA assays, dilution is done to optimize the signal-to-noise ratio and to reduce the background signal.
ELISA refers to a type of immunoassay that uses antibodies to dete… #
Related terms include immunological assay and biochemical assay, which describe the type of assay and the type of reaction used to detect the analyte, respectively. In ELISA assays, the specificity and sensitivity of the antibodies are critical to the success of the assay.
Enzyme refers to a protein that catalyzes a chemical reaction, whi… #
Related terms include catalyst and biocatalyst, which describe a substance that speeds up a chemical reaction and a biological molecule that speeds up a chemical reaction, respectively. In ELISA assays, enzymes are used to detect the analyte, and their activity and stability can affect the results.
Epitope refers to the specific region on an antigen that is recogn… #
Related terms include antigenic determinant and binding site, which describe the region on an antigen that is recognized by an antibody and the region on an antibody that binds to an antigen, respectively. In ELISA assays, the epitope is the target of the antibody, and its structure and conformation can affect the results.
Fluorescence refers to the emission of light by a molecule after <… #
Related terms include phosphorescence and bioluminescence, which describe the emission of light by a molecule after absorption of light and the emission of light by living organisms, respectively. In ELISA assays, fluorescence is used to detect the analyte, and its sensitivity and specificity are critical to the success of the assay.
Hapten refers to a small molecule that can trigger an immune respo… #
Related terms include antigenic determinant and epitope, which describe the region on an antigen that is recognized by an antibody and the specific region on an antigen that is recognized by an antibody, respectively. In ELISA assays, haptens are used to detect the analyte, and their structure and conformation can affect the results.
Immobilization refers to the process of attaching a molecule to a… #
Related terms include coating and adsorption, which describe the process of attaching a molecule to a surface and the process of attracting and holding molecules to a surface, respectively. In ELISA assays, immobilization is done to capture the analyte, and the concentration and purity of the immobilized molecule can affect the results.
Immunoglobulin refers to a type of protein that is produced by the immune… #
Related terms include antibody and immunoglobulin G, which describe a protein that is produced by the immune system and a type of immunoglobulin, respectively. In ELISA assays, immunoglobulins are used to detect the analyte, and their specificity and sensitivity are critical to the success of the assay.
Incubation refers to the process of allowing a reaction to occur,… #
Related terms include reaction time and temperature control, which describe the time allowed for a reaction to occur and the control of temperature to optimize the reaction, respectively. In ELISA assays, incubation is done to allow the binding of the antibody to the analyte, and the duration and temperature of incubation can affect the results.
Inhibition refers to the process of reducing or blocking a chemica… #
Related terms include competitive inhibition and non-competitive inhibition, which describe the inhibition of a reaction by a competing molecule and the inhibition of a reaction by a molecule that binds to a different site, respectively. In ELISA assays, inhibition is used to detect the analyte, and its sensitivity and specificity are critical to the success of the assay.
Interference refers to the presence of a substance that can affect … #
Related terms include background signal and noise, which describe the signal that is not related to the analyte and the random fluctuations in the signal, respectively. In ELISA assays, interference can be caused by contaminants or inhibitors, and its effect can be minimized by using controls and optimizing the assay conditions.
Label refers to a molecule that is attached to a detection reagent … #
Related terms include tag and probe, which describe a molecule that is attached to a detection reagent and a molecule that is used to detect a target molecule, respectively. In ELISA assays, labels are used to detect the analyte, and their sensitivity and specificity are critical to the success of the assay.
Microplate refers to a plate with multiple wells that is used to p… #
Related terms include well and plate reader, which describe a single well on a microplate and an instrument that is used to measure the signal from multiple wells, respectively. In ELISA assays, microplates are used to perform multiple assays simultaneously, and their quality and consistency can affect the results.
Optimization refers to the process of adjusting the conditions of… #
Related terms include validation and verification, which describe the process of evaluating the performance of an assay and the process of confirming the results of an assay, respectively. In ELISA assays, optimization is done to maximize the signal-to-noise ratio and to minimize the background signal.
Plate reader refers to an instrument that is used to measure the s… #
Related terms include microplate reader and spectrophotometer, which describe an instrument that is used to measure the signal from multiple wells and an instrument that is used to measure the interaction between light and a sample, respectively. In ELISA assays, plate readers are used to measure the signal from multiple wells, and their accuracy and reliability are critical to the success of the assay.
Precision refers to the degree of consistency of a measurement, wh… #
Related terms include accuracy and reliability, which describe the degree of closeness of a measurement to the true value and the ability of a method to produce consistent results, respectively. In ELISA assays, precision is critical, as small errors can lead to incorrect conclusions.
Quality control refers to the process of ensuring that an ELISA as… #
Related terms include quality assurance and validation, which describe the process of ensuring that an assay is performed correctly and the process of evaluating the performance of an assay, respectively. In ELISA assays, quality control is done by using controls and standards to validate the results.
Reagent refers to a substance that is used in an ELISA assay to <i… #
Related terms include antibody and enzyme, which describe a protein that is used to detect the analyte and a protein that is used to catalyze a chemical reaction, respectively. In ELISA assays, reagents are used to detect the analyte, and their quality and purity can affect the results.
Sample preparation refers to the process of preparing a sample for an … #
Related terms include extraction and purification, which describe the process of isolating a substance from a sample and the process of removing impurities from a sample, respectively. In ELISA assays, sample preparation is done to concentrate the analyte and to remove impurities that can interfere with the assay.
Sensitivity refers to the ability of an ELISA assay to detect</… #
Related terms include limit of detection and limit of quantitation, which describe the smallest amount of analyte that can be detected and the smallest amount of analyte that can be quantitated, respectively. In ELISA assays, sensitivity is critical, as small errors can lead to incorrect conclusions.
Specificity refers to the ability of an ELISA assay to detect</… #
Related terms include cross-reactivity and interference, which describe the reaction of an antibody with a molecule that is not the analyte and the presence of a substance that can affect the results of the assay, respectively. In ELISA assays, specificity is critical, as small errors can lead to incorrect conclusions.
Standard refers to a sample with a known concentration of analyte,… #
Related terms include calibrator and control, which describe a sample with a known concentration of analyte and a sample with a known concentration of analyte that is used to validate the results, respectively. In ELISA assays, standards are used to ensure accuracy and reliability, and their concentration and purity can affect the results.
Substrate refers to a substance that is used in an ELISA assay as… #
Related terms include reagent and co-substrate, which describe a substance that is used in an assay and a substance that is used in conjunction with a substrate, respectively. In ELISA assays, substrates are used to detect the analyte, and their quality and purity can affect the results.
Validation refers to the process of evaluating the performance of… #
Related terms include verification and qualification, which describe the process of confirming the results of an assay and the process of verifying the performance of an instrument, respectively. In ELISA assays, validation is done to ensure that the assay is specific and sensitive enough to detect the analyte.
Wash refers to the process of removing unbound reagents from a sam… #
Related terms include rinsing and aspiration, which describe the process of removing liquid from a sample and the process of removing liquid from a sample using a pipette, respectively. In ELISA assays, washing is done to remove impurities and to reduce the background signal.