Thermo Scientific Pierce
Super Signal West Dura Extended Duration Substrate
34076
Info: View Product View Product Specs
The experiment was designed to quantify low abundance proteins (membrane receptors) in mammalian cell lines using western blot technique. 40 micrograms of cell lysate was resolved in SDS-PAGE, western transferred in a nitrocellulose membrane and probed with specific antibody. Blot was either treated with Super Signal West Dura ECL substrate or normal ECL substrate for 2.5 minutes and x-Ray film was developed.
Visualization of specific HRP activity in western blot
RIPA buffer, SDS-PAGE, Western transfer unit, nitrocellulose membrane, primary and HRP conjugated secondary antibody, X-ray films, developer, and fixer
Cells were treated with growth factor and proteins were isolated with RIPA buffer, Equal amounts of proteins were resolved in 10% SDS-PAGE and western transferred in a nitrocellulose membrane. Nitrocellulose membranes were generated in duplicate. Membranes were treated with specific primary (1:1000) and anti-species HRP conjugated secondary antibodies. One membrane was treated with normal ECL substrate from GE healthcare and another with Super Signal West Dura Extended Duration Substrate. HRP activity was captured on x-ray film.
None
Membrane developed in normal ECL substrate from GE healthcare showed weak or no signal for low abundance proteins and vanished after 20 mins of treatment. On the other hand, membrane developed with Super Signal West Dura Extended Duration Substrate from Thermo Fisher showed intense signal which was detected for at least 2.5 hrs of the treatment.
Got intense signal for a long period
Some non-specific signals with dusty background were detected
Good performence
Product Name: SuperSignal West Dura Extended Duration SubstrateSupplier Page