Images are generated utilizing time resolved fluorescence (TRF) mode detection of Europium (Eu) which has a 1 ms fluorescence lifetime. Detection with ScanLater TRF Western Blot Detection Cartridge (4). Eu-labeled ScanLater Secondary Antibody (3) binds to primary antibody. Use existing primary antibody (1) for binding to protein of interest (2). Membranes are incubated with Europium-chelate labeled secondary antibodies or streptavidin that bind specifically to the primary antibody bound to the protein of interest (Figure 1).įigure 1. ScanLater ® Western Blot System workflow follows standard gel loading and blotting methods up to the secondary antibody incubation step. In addition, a comparison of Scanlater Western Blot System to a chemiluminescence method demonstrates improved sensitivity. We demonstrate extended dynamic range due to reduced background as well as stability of detection over time and number of reads. Here we report on a novel system for western blot membrane protein analysis that is incorporated into SpectraMax ® i3 and SpectraMax ® Paradigm ® Multi-Mode Microplate Readers. However, each technique has its limitations, and there is a continuing need to improve quantitation, accuracy, and dynamic range. Various techniques are used to detect proteins on western blot membranes including fluorescence and chemiluminescence. Protein detection is important for pharmaceutical and clinical research today, and western blots are among the most common methods employed for this purpose.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |