NeoPRO Nano Western ECL Substrate

The NeoPRO Nano Western ECL Substrate is a two-component chemiluminescent substrate designed for Western blotting and HRP-based protein detection. It provides a mid-picogram detection level and is intended for routine Western blot applications where reliable chemiluminescent detection of protein targets is required.

The substrate is based on a luminol-derived chemiluminescent detection system. Peroxidase-catalyzed oxidation generates light, while reaction components incorporated into the formulation increase signal intensity and duration. This enables detection of proteins immobilized on Western blot membranes following antibody incubation.

The NeoPRO Nano Western ECL Substrate is supplied as a two-solution kit containing a luminol derivative/enhancer solution and a peroxide solution. The two solutions are mixed at a 1: 1 ratio immediately before use to prepare the working solution.

The substrate can be used with standard Western blotting workflows involving SDS-PAGE, membrane transfer, blocking and HRP-conjugated antibody detection. The recommended antibody dilution ranges and controlled chemiluminescent detection procedure provide a practical starting point for mid-picogram protein detection.

• Two-component chemiluminescent substrate for Western blotting
• Designed for mid-picogram protein detection
• HRP-dependent chemiluminescent detection
• Luminol derivative/enhancer and peroxide solutions supplied separately
• Working solution prepared by mixing the two components at a 1: 1 ratio
• Suitable for routine Western blot applications
• Compatible with HRP-conjugated secondary antibodies
• Suitable for chemiluminescent signal acquisition using imaging devices or autoradiography film
• Recommended primary antibody dilution: 1: 100–1: 5,000
• Recommended secondary antibody dilution: 1: 1,000–1: 15,000
• Research use only

1. Prepare the working solution: Mix Solution A and Solution B at a 1: 1 ratio immediately before use.
2. Prepare the membrane: Following primary and HRP-conjugated secondary antibody incubation, rinse the membrane twice with TBS-T.
3. Apply the substrate: Use approximately 0.1 mL of NeoPRO Nano working solution per cm² of membrane.
4. Incubate: Ensure that the entire membrane surface is covered and incubate for approximately 1.5 minutes.
5. Acquire the signal: Detect the chemiluminescent signal using an imaging device or autoradiography film.
6. Optimize exposure: For an unknown signal, initial exposure times of 15 seconds, 30 seconds, 1 minute and 5 minutes can be tested.