UltraScence Pico Plus Western Substrate
The UltraScence Pico Plus Western Substrate, as a luminol-based enhanced chemiluminescent substrate, is sensitive and compatible with conducting immunoblots with horseradish peroxidase (HRP) – conjugated secondary antibodies. The low picogram or high femtogram detection of antigen is enabled by UltraScence Pico Plus Western Substrate’s excellent sensitivity and long signal duration. Further, its long chemiluminescent signal duration makes both digital and film-based imaging possible without any loss of the signal. Appropriate primary and secondary antibody dilutions are suggested for attaining optimal signal intensity and duration.
- No optimization required, switching to the UltraScence Pico Western Substrate from other brands, such as Thermo Scientific™ SuperSignal™ Pico Plus and Bio-Rad™ Clarity™, does not require optimization or protocol changes.
- High degree of sensitivity and enhanced chemiluminescence duration, UltraScence Pico Plus Western Substrate enables an accurate low picogram or high femtogram detection of protein on the same immunoblot after a single exposure.
- Optimized for use with PVDF and nitrocellulose membranes.
- Compatible with Western Blotting Markers.
- Optimized for film- and CCD-based imaging.
- Keep the membrane moist in the wash buffer while preparing the substrate mixture. Make sure the membrane does not dry out in the next steps.
The chemiluminescent substrate solution is sufficiently stirred to prepare the 0.1ml of solution / cm2 of the membrane.
- For a small membrane (7 x 8.5 cm), 4 ml of solution is sufficient.
- For a medium-sized membrane (8.5 x 135 cm), 10 ml of solution is sufficient.
- Place the membrane on a clean, flat surface or in a clean container.
- Remove the membrane from the chemiluminescent substrate solution and drain the excess solution.
- Place the membrane in a plastic sheet protector or plastic wrap to prevent the film from drying out.
- Imaging the membrane with a digital imager or by exposure to the X-ray film.
The membrane was probed with a 1:10000 dilution of β-actin antibody (GTX109639) and then serially diluted HeLa whole cell lysates were prepared using a rabbit IgG IgG HRP conjugated secondary antibody (GTX213110-01, 1:10,000). , and apply in electrophoresis and protein transfer. The same blot was incubated with 5 ml of UltraScence Pico Plus Western substrate (CCH321-B100ML). Blots were simultaneously exposed for 10 seconds, 30 seconds and 60 seconds using a UVP BIO SPECTRUM-AC digital imaging system.
* SuperSignal West Pico PlUS is a registered trademark of Thermo Fisher Scientific. The trademark holder is not affiliated with Bio-HeliX Co., Ltd. and does not recognize this product.