ECL chemiluminescence Kit is a high sensitivity chemiluminescence kit, the basic principle is based on luminol ECL chemiluminescence. The product can react with the horseradish peroxidase (HRP) conjugated on the secondary antibody to produce fluorescence, which can be detected by X-ray pressing or other appropriate fluorescence imaging equipment (CCD camera, etc.).

 

The company currently produces two kinds of ECL chemiluminescence kits, namely G2014 ECL chemiluminescence kit and G2020 ultra-sensitive ECL chemiluminescence kit. For the detection of target proteins with high abundance, such as internal reference proteins, it is recommended to use G2014, which can detect target proteins with a content greater than or equal to 2 ng. However, for target proteins with low abundance that are difficult to detect, G2020 is recommended. Compared with G2014, the sensitivity of this product is at least 5 times higher, and pg level of protein can be detected, and the luminescence is more stable.

 

 

Storage and transportation

 

Wet ice transportation; Stored at 4℃, away from light, valid for 12 months. If it is not used for a long time, it can be stored at -20℃ for a longer time.

 

steps

 

1. Prepare ECL working solution: mix ECL A solution and ECL B solution in equal volume and store at 4℃ under dark conditions. Available now for use within 2 days.

 

2. In the Western experiment, PVDF membrane was incubated by the second antibody, washed several times, and the excess liquid was sucked out by filter paper. Paste two layers of PE gloves or other transparent films on the exposure box, place the PVDF membrane protein face up between the two layers of the exposure box, add the mixed ECL working solution and cover the film for 1-2 min.

 

3. Use filter paper or absorbent paper to absorb the ECL working fluid, cover the upper film and begin to press the film.

 

4. Develop and fix the finished film with developing and fixing reagents (G2019, G2023, G2024 are recommended). Adjust exposure conditions according to luminescence intensity.

 

Matters needing attention

 

1. The suction head of ECL liquid A and liquid B must be replaced during the suction process. Mutual contamination of liquid A and liquid B will lead to gradual failure of liquid A or liquid B. Keep sealed after use.

 

2. If the background is very deep after exposure, the possible reason is that the concentration of the secondary antibody is too high, or the concentration of the primary antibody is too high, or the sealing fluid is not suitable, so change to other sealing fluid.

 

3. If the fluorescence quenching is rapid, the possible cause is that the fluorescence of the destination strip is too strong, causing HRP to rapidly consume ECL.

 

4. If there is no luminescence signal, the expression of the target protein may be weak, which may prolong the tablet pressing time.

 

5. Please wear lab coat and disposable gloves during operation

 

 

This product is only for scientific research purposes, not for clinical diagnosis!