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G2014-50ML 2x50ml ECL Chemiluminescence Kit for Protein Detection

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G2014-50ML 2x50ml ECL Chemiluminescence Kit for Protein Detection

The ECL chemiluminescence kit is a highly sensitive chemiluminescence kit based on the principle of Luminol ECL chemiluminescence. This product can chemically react with horseradish peroxidase (HRP) coupled with secondary antibodies to produce fluorescence, and the sample can be detected by X-ray tablet pressing or other appropriate fluorescence imaging equipment (CCD camera, etc.).

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

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Technical Specification

Storage and Handling Conditions

Wet ice bag transportation; Stored at 4ºC in the dark; valid for 12 months.
 

Preparation for the experiment

Protease inhibitor is provided. The RIPA lysate (strong) should be added with protease inhibitors before use. G2006, G2007, G2008, etc., are recommended to prevent protein degradation. The RIPA lysate (strong) mentioned in the following usage means that protease inhibitors have been added.

Assay Protocol / Procedures

  1. Preparation of ECL working fluid: Mix ECL solution A and ECL solution B in equal volume, and keep them at 4ºC away from light. Use it right after it is ready, within 2 days.
  2. In the Western blot experiment, PVDF membrane was incubated by the secondary antibody, washed for several times, and the excess liquid was absorbed by the filter paper. Two layers of PE gloves or other transparent films were affixed to the exposure box. The PVDF membrane protein was placed face up between the two layers of the exposure box. The mixed ECL working solution was added to cover the film and placed on the film for 1-2 min.
  3. Use filter paper or absorbent paper to absorb ECL working solution, cover the upper film and start pressing film.
  4. Development and fixing reagents (G2019, G2023 and G2024 are recommended) shall be used to develop and fix the pressed film. Adjust the exposure conditions according to the luminous intensity.

 

Notes:

  1. The pipette tips must be replaced during the liquid transferring process of ECL liquid A and liquid B. Cross contamination of liquid A and liquid B will lead to the gradual failure of liquid A or B. In addition, the contamination of metal ions will reduce the sensitivity of this reagent. Please pay attention to use clean pipette tips, Seal well after use.
  2. If the background after exposure is very deep, the reason may be that the concentration of the secondary antibody is too high, or the concentration of the primary antibody is too high, or the sealing solution is not suitable, and other sealing solution should be used.
  3. If the fluorescence quenches rapidly, it may be due to the over-strong fluorescence of the target band, resulting in the rapid consumption of ECL by HRP.
  4. If there is no luminous signal, it may be that the target protein expression is very weak, which can prolong the tablet pressing time.
  5. Please wear a lab coat and disposable gloves during operation
     

For research only!

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