Femto Plus Western Substrate

DATA SHEET

Femto Plus  Western Substrate is a highly sensitive and compatible chemiluminescent substrate that can be used for  immunoblots with horseradish peroxidase (HRP) – conjugated secondary antibodies.  This luminol-based enhanced chemiluminescent substrate has excellent sensitivity and long signal duration, hence the Pico Plus Western Substrate allows for  the detection of antigens in the low femtogram range. Additionally, its long-lasting chemiluminescent signal allows for both digital and film-based imaging without any loss of signal. To achieve optimal signal intensity and duration, we recommend using appropriate primary and secondary antibody dilutions.

• No optimization required
• Compatible with PVDF and nitrocellulose membranes

material needed but not provided

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

Reaction Set up

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.