ELISA kit method is a widely used and well proven solid-phase immunoassay technique which can detect or quantify one or several different analytes in an aqueous sample. It is useful in helping provide accurate, sensitive and consistent results, where each target protein is researched and each kit is calibrated to provide physiologically relevant sensitivity.
FEATURES AND BENEFITS OF OUR ELISA KIT
- Broad range of targets available over 2,000 at the moment and we are constantly adding more.
- Optimised for accurate, sensitive and consistent performance. They are put through rigorous quality control specifications and are manufactured in ISO accredited facilities in order to help ensure excellent quality and reproducibility.
- Validated for sample type and for species specificity, thereby providing a physiologically relevant sensitivity.
- Contains all the reagents and solutions required to perform the ELISA experiment including the protein standards and controls.
- It is flexible and can be adapted to be used on an automated equipment in order for higher throughput systems.
- A detailed protocol insert with an easy step by step instructions guide is provided.
TYPICAL CONTENTS OF AN ELISA KIT
- Antibody-coated 96-well plate or 96-well pre-coated strip plate (12×8). Usually a polystyrene solid support which can be stored for 6-10 months at room temperature (2-8°C).
- Standards and controls. Provided either in a pre-diluted format or may require some dilution. These need to be included with each plate so the results are valid and meaningful.
- Primary detection antibody (typically biotinylated) and a secondary detection reagent (usually HRP-streptavidin). Key component of the assay, a highly purified IgG detection antibody is recommended and the preferred enzyme reagent used is either horseradish peroxidase (HRP) or alkaline phosphatase (AP).
- Sample diluent buffers and wash buffer concentrate (20x). Need to bring both these reagents to room temperature before using them.
- Substrate and stop solution. Substrate solution needs to be kept in the dark until required for use, since any expose to light could affect its chemical activity. The stop solution needs to be bought to room temperature before use, it also needs to be clear without any crystals present.
- Plate cover.
- Instruction manual.
AN OVERVIEW OF THE ELISA KIT METHOD
- A typical sandwich ELISA method employ specific capture antibodies that have been pre-coated on a 96-well plate.
- Standards and samples are then pipetted into each of the specified wells, this will result in the target protein in the standards and samples binding to the immobilized antibody.
- The wells are then thoroughly washed (usually using PBS) in order to remove any non-specific binding, followed by the addition of the biotin-labelled detection antibody.
- After a second washing step to remove away any unbound biotinylated antibodies, AP or HRP-conjugated streptavidin is pipetted into each well, followed by a colorimetric substrate solution. Finally a stop solution is added to each well in order to end the reaction.
- A standard curve is prepared using the data obtained from the standards and this is used to interpolate the concentration of the samples.
- The intensity of colour that develops in each of the wells is proportional to the amount of target protein bound.