Protocol for

Capturing Multiplex Chemiluminescent ELISAs

Link to Full Protocol: http://www.carestreamhealth.com/capturing-multiplex-AP0087-s ...

Abstract or Description:

CCD cameras are widely used to capture chemiluminescent images of Western blots and ELISAs (enzyme-linked immunosorbent assay). ELISAs are used as a diagnostic tool to determine the concentration of a specific protein in a sample via antigen-antibody binding. In most cases, the sample or antigen is bound to the bottom of a 96-well plate, a specific antibody linked to an enzyme is incubated over the antigen, and then a substance that cleaves to the enzyme is added and gives a colorimetric or chemiluminescent signal.

Sandwich ELISAs consist of antibodies pre-coated onto a 96-well plate, followed by incubation with a protein sample containing unknown concentrations of antigen. The plate is then incubated with a detection antibody, which is linked to an enzyme that is cleaved when a chromogen or substrate is added to the plate and produces a colorimetric, chemiluminescent, or fluorescent signal.

A multiplex sandwich ELISA was developed by Quansys Biosciences (Logan, UT), where several different antibodies are bound to the bottom of each well of a 96-well plate in a grid format. After cell lysate and detection antibody hybridizations are complete, the chemiluminescent substrate is added, and the plate is immediately imaged with a CCD camera. The 4 mega-pixel CCD camera in the Carestream Image Station 4000MM (IS4000MM) digital imaging system is capable of imaging these small antibody grids with excellent resolution and signal intensity. The following report presents an evaluation of the Q-Plex™ 9-plex Calibration and 16-plex Human Cytokine Screen kits.

Citation:

Attachments: No Attachments

Posted By: Carestream Health on 1/7/2011 12:22:22 PM

Comments

Add Comment