UltraScence Femto Western Substrate
Description
The UltraScence Femto Western Substrate, as a luminol-based enhanced chemiluminescent substrate, is sensitive and compatible with conducting immunoblots with horseradish peroxidase (HRP) – conjugated secondary antibodies. The mid femtogram to low femtogram detection of antigen is enabled by UltraScence Femto 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.
Features
- No optimization required, switching to theUltraScence Femto Western Substrate from other brands, such as Thermo Scientific™ SuperSignal™ West Femto and Wako™ ImmunoStar™ LD, does not require optimization or protocol changes.
- High degree of sensitivity and enhanced chemiluminescence duration, ultraScence Femto Western Substrate enables an accurate mid picogram to low 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.
- Patent pending product.
Chemiluminescent Development
- 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.
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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.
Performance
Membranes were probed with Rabbit anti β-actin antibody (GTX109639) diluted at 1:5,000 of and then with Goat anti Rabbit HRP-conjugated secondary antibody (GTX213110-01, 1:10,000) after serial dilution mock RD cell lysates were prepared and applied in electrophoresis and protein transfer. Identical blots were incubated with 400 μL of UltraScence Pico Ultra Western substrate (CCH345) and UltraScence Femto Ultra Western substrate (CCH365). The blots were simultaneously exposed for 10 seconds, 30 seconds and 60 seconds a using Chemlux SPX-600 Series digital imaging system.
* ImmunoStar™ LD is a registered trademark of FUJIFILM™ Wako™ Pure Chemical Corporation and Trident™ femto Western HRP Substrate is a registered trademark of GeneTex™. The trademark holder is not affiliated with Bio-HeliX Co., Ltd. and does not endorse these products.
ELISA Use
Comparison
