The OneStep® ELISA Kit is a specialized format of the traditional Enzyme-Linked Immunosorbent Assay (ELISA) designed to streamline the assay process, reducing the number of steps and time required to obtain results. This "one-step" approach typically combines the capture and detection steps into a single process, enhancing ease of use and efficiency.

Key Components and Technical Details

Principle of the Assay

• The OneStep® ELISA operates on a simplified principle where the process of capturing the target analyte and detecting it are combined into a single step, reducing procedural complexity and time.

Kit Components

• Pre-coated Microplate: A 96-well plate or another format pre-coated with capture antibodies specific to the target analyte.
• Detection Reagent: Often a conjugate containing both capture and detection antibodies or a single reagent that facilitates both binding and signal generation. This might include enzyme-conjugated antibodies or a reagent that binds directly to the target analyte.
• Substrate Solution: A reagent that reacts with the enzyme linked to the detection antibody to produce a colorimetric or chemiluminescent signal proportional to the amount of target analyte.
• Stop Solution: Used to halt the enzymatic reaction and stabilize the signal for measurement.
• Washing Buffer: Typically included to wash away unbound substances and reduce background interference.

Sample Preparation

• Homogenates/Lysates: For tissue or cell lysates, proper preparation ensures effective extraction of the target analyte.
• Serum/Plasma: For blood-based assays, samples need to be handled and processed to maintain analyte stability and avoid degradation.

Assay Procedure

• Incubation: Samples, standards, and the detection reagent are added to the pre-coated wells simultaneously. This step combines the capture and detection processes.
• Washing: Following incubation, the wells are washed to remove unbound substances, if necessary.
• Substrate Reaction: The substrate solution is added to develop the colorimetric or chemiluminescent signal.
• Stop Reaction: A stop solution is added to halt the enzymatic reaction and stabilize the signal.
• Reading: Optical density or fluorescence/intensity is measured using a microplate reader or a similar device.

Quantification

• A standard curve generated from known concentrations of the target analyte is used to determine the concentration in the samples based on the measured signal.

Applications

• Clinical Diagnostics: For rapid assessment of biomarkers in patient samples.
• Research: Facilitating high-throughput analysis of proteins, hormones, or other analytes in various research applications.
• Quality Control: Used in manufacturing and quality control processes to measure analyte levels efficiently.