Description
Ready-to-use Pico ECL substrate for Western blot
GenieBlot Pico ECL Ready-to-Use Substrate is a premixed, single-bottle ECL HRP substrate for routine Western blot detection at picogram sensitivity, with no mixing step required. It delivers bright, durable chemiluminescent signal for fast, reproducible imaging.
This sensitive, luminol-based enhanced chemiluminescent substrate detects horseradish peroxidase (HRP) on immunoblots. Unlike conventional two-part ECL kits, there is no need to mix substrate and peroxide in equal volumes before use, which avoids pipetting errors and contamination and improves consistency between experiments. It detects low-picogram levels of protein and gives prolonged chemiluminescence, allowing imaging for up to 5 hours. Use it with HRP-labelled antibodies or nucleic acid probes on NC or PVDF membranes, for Western blot, EMSA and other blot detection, imaged on X-ray film or a CCD imaging system.
For low-abundance targets, use the GenieBlot Femto ECL Ready-to-Use Substrate. For research use only.
Key Features
| Easy to use | Ready-to-use single-component format with no mixing, minimising pipetting errors and contamination. | |
| High sensitivity | Detects proteins at low picogram levels. | |
| Long-lasting signal | Prolonged chemiluminescence allows imaging for up to 5 hours. | |
| Flexible imaging | Compatible with X-ray film and CCD imaging systems. |
Specifications
| Technology | Luminol-based enhanced chemiluminescence (HRP substrate) |
| Format | Single-component, ready-to-use working solution |
| Sensitivity | Low picogram protein detection |
| Signal duration | Imaging possible for up to 5 hours |
| Applications | Western blot, EMSA and other blots using HRP-labelled antibodies or nucleic acid probes |
| Membranes / detection | NC or PVDF; X-ray film or CCD imager |
| Recommended volume | About 70 μl/cm2 membrane |
| Shelf life | 2 years |
Components
| Components | 100 ml | 500 ml |
| GenieBlot Pico ECL Ready-to-Use Substrate | 100 ml | 2 × 250 ml |
Storage
Store at 2 ~ 8°C.
- Wash the membrane
- Perform the Western blot as required, then wash off the secondary antibody with PBST/PBS or TBST/TBS.
- Apply the substrate
- Lift the membrane with flat tweezers and drain the wash buffer on filter paper without letting the membrane dry completely.
- Pipette the substrate evenly over the membrane to cover it completely (about 70 μl/cm2, adjustable to preference) and incubate at room temperature for 1 min.
- Drain excess substrate
- Hold the membrane vertically on filter paper with flat tweezers for a few seconds to remove excess solution.
- Image the blot
- X-ray film: place the membrane quickly between two layers of plastic wrap, pressing out any bubbles. Place it protein side up in the film cassette and secure the edges with tape. Choose an exposure time based on signal development, or take several exposures of different lengths.
- CCD imager: no plastic wrap is needed; image according to the instrument's instructions.
Q1: Do I need to mix two reagents before use?
- No. The substrate is a single-component, ready-to-use solution, so there is no need to mix substrate and peroxide in equal volumes. This avoids pipetting errors and contamination and improves reproducibility.
Q2: What antibody dilutions should I use?
- Because the substrate is highly sensitive, dilute primary antibodies 1:1,000–1:4,000 and secondary antibodies 1:5,000–1:10,000.
Q3: How can I reduce high background?
- Use high-quality plastic wrap, as impurities can contaminate the membrane, raise background or quench the signal; shorten the exposure time, as overexposure darkens the background; and check that antibody dilutions are within the recommended range.
Q4: Why is my signal weak?
- Possible causes include underexposure, a low-abundance target or HRP inhibition. Increase the exposure time, avoid sodium azide in buffers, and for low-abundance targets use the GenieBlot Femto ECL Ready-to-Use Substrate.
