The UltraScence Pico Ultra 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 to mid femtogram detection of antigen is enabled by UltraScence Pico Ultra 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.

Product Name: UltraScence Pico Ultra Western Substrate
Product Item No.: CCH345-B100ML
Dimensions: 100ML
Substrate Type: HRP (Horseradish Peroxidase) Substrate
Contents/Storage: Store at R.T. for 1 year, 4°C for 2 years

- No optimization required. 
Switching to theUltraScence Pico Ultra Western Substrate from other brands, such as Thermo Scientific™ SuperSignal™ West Dura and Millipore™ Immobilon™ substrates, does not require optimization or protocol changes.

- High degree of sensitivity and enhanced chemiluminescence duration. 
UltraScence Pico Ultra Western Substrate enables an accurate low picogram to mid 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.

1. 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.

2. 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.

3. Place the membrane on a clean, flat surface or in a clean container.

4. Remove the membrane from the chemiluminescent substrate solution and drain the excess solution.

5. Place the membrane in a plastic sheet protector or plastic wrap to prevent the film from drying out.

6. Imaging the membrane with a digital imager or by exposure to the X-ray film. 

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 4 ml of UltraScence Pico Ultra Western ECL substrate (CCH345). The blots were simultaneously exposed for 1 minute, 5 minutes and 10 minutes using a Chemlux SPX-600 Series digital imaging system. 
*Immobilon™ Western Chemiluminescent HRP Substrate is a registered trademark of Millipore and Amersham ECL™ Prime Western Blotting Detection Reagent is a registered trademark of GE Healthcare. The trademark holder is not affiliated with Bio-HeliX Co., Ltd. and does not endorse these products.