UltraScence Femto Plus Western Substrate

The UltraScence Femto 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 UltraScence Femto Plus Western Substrate is designed for the detection of the target proteins in amounts that are too small to be seen with typical ECL substrates. The low femtogram detection (mid-atto) of antigen is enabled by UltraScence Femto 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. Moreover, the high stability of UltraScence Femto Plus Western Substrate makes a 2-year- storage at room temperature possible, without any compromise on the performance.

• No optimization required. Switching to the UltraScence Pico Plus Western ECL Substrate from other brands, such as Pierce ECL and GE Healthcare, does not require optimization or protocol changes.
• High degree of sensitivity and enhanced chemiluminescence duration. UltraScence Pico Plus Western ECL Substrate enables an accurate low picogram to 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.

• Stable for up to 24 months at 4 °C, do not freeze it.
• Ship at 4 °C, beware of shipping in any condition beneath 0 °C.

1. Keep the membrane moist in the wash buffer while preparing the substrate mixture. Please ensure the membrane does not dry out during the subsequent steps.
2. Mix Luminol solution and Peroxide Solution in a 1:1 ratio, and thoroughly agitate the chemiluminescent substrate solution well for preparing the 0.1 ml of solution / cm² of membrane.
- For a mini-sized membrane (7 x 8.5 cm), 4 ml of solution is sufficient.
- For a midi-sized membrane (8.5 x 13.5 cm), 10 ml of solution is sufficient.
3. Place the membrane with the protein side up on a clear and level surface or in a clean container.
4. Remove the membrane from the chemiluminescent substrate solution and drain off excessive solution.
5. Place the membrane in a plastic sheet protector or in plastic wrap to prevent the membrane from drying.
6. Image the membrane with a digital imager or by exposing to the X-ray film.

• A compatible Chemiluminescence or X-ray Imaging Systems.
• A plastic sheet protector or plastic wrap to prevent the membrane from drying.