Product Information

Product Description/Introduction

ECL chemiluminescence Kit is a highly sensitive chemiluminescence kit based on luminol ECL chemiluminescence. This product can react with horseradish peroxidase (HRP) conjugated on secondary antibody to produce fluorescence and detect samples by using X-ray lithography or other appropriate fluorescence imaging equipment (CCD camera, etc.). On the basis of G2014, it is further optimized with higher sensitivity and more stable color development, which is suitable for the detection of low abundance target proteins.

The company currently produces two kinds of ECL chemiluminescence kits, namely G2014 ECL chemiluminescence Kit and G2020 Hypersensitive ECL chemiluminescence Kit. For the detection of target proteins with high abundance, such as internal reference proteins, G2014 is recommended, and the detectable content of target proteins is greater than or equal to 2 ng. For low abundance and difficult to detect target proteins, G2020 is recommended. The sensitivity of this product is 5 times higher than that of G2014, and it can detect pg level proteins, and the luminescence is more stable.

Storage and Shipping Conditions

Ship with wet ice; Store in the dark at 4℃, valid for 12 months. If not used for a long time, it can be stored at -20℃.

Product Components

Assay Protocol/Procedures

1. Preparation of hypersensitive ECL working solution: Mix the hypersensitive ECL A solution and hypersensitive ECL B solution in equal volumes, and store in the dark at 4℃. Use it within 2 days.

2. In Western experiment, PVDF membrane is incubated with secondary antibody, washed several times, and excess liquid is absorbed by filter paper. Stick two layers of PE gloves or other transparent films on the exposure box, place the protein side of the PVDF film between the two layers of the exposure box, add the mixed ECL working solution to cover the film and place it on the film for 1-2 min.

3.  Remove the ECL working liquid with filter paper or absorbent paper, cover the upper film and start pressing the film.

4.  The pressed film should be developed and fixed with developing and fixing reagents (G2019, G2023, G2024 are recommended). Adjust exposure conditions according to luminous intensity.

Note

1. High sensitive ECL A liquid and high sensitive ECL B liquid in the suction process, the suction head must be replaced. The mutual contamination of liquid A and liquid B will lead to the gradual failure of liquid A or liquid B. In addition, the contamination of metal ions will reduce the sensitivity of this reagent, please pay attention to the use of clean suction head. Store sealed after use.

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

3. If the fluorescence is quenched quickly, the possible cause is that the fluorescence of the target band is too strong, resulting in rapid consumption of ECL by HRP.

4. If there is no luminescence signal, the target protein may be weakly expressed and the compression time may be extended.

5. For your safety and health, please wear safety glasses, gloves, or protective clothing.

For Research Use Only!