UltraScence Femto Plus Western Substrate Powder

The UltraScence Femto Plus Western Substrate Powder, 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 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.

• Ready-to-ship and significantly longer shelf life in the powder form!
• Significant reductions in transportation and storage costs, resulting in less carbon footprint!
• Empower custom production anywhere in the world!
• Significantly more sensitive than leading manufacturers on the market

• Prepare two 5 L bottles. One amber bottle is for luminol solution, and other normal bottle is for peroxide solution. (Make sure the bottles are acid and alkali resistant.)
• Unpack the bag and pour all the powder in bottle.
• Rinse residual powder in bag with deionized water.
• Add deionized water to 5 L.

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.

Membranes were probed with GFP tag Rabbit PolyAb diluted at 1:10,000 of and then with Goat Anti-rabbit IgG/HRP secondary antibody (1:10,000) after serial dilution EGFP (Enhanced Green Fluorescent Protein) were prepared and applied in electrophoresis and protein transfer. Identical blots were incubated with 400 μL of UltraScence Femto Plus Western substrate (CCH375). The blots were simultaneously exposed for 5 seconds, 10 seconds, 30 seconds, 60 seconds and 180 seconds and using Chemlux SPX-600 Series digital imaging system. The comparison data of the UltraScence Femto Plus Western Substrate between the liquid form and powder form is shown below.