ECL Luminescence Reagent #abs920

Description

Western blot substrate luminescence detection reagent can be catalyzed by the horseradish peroxidase labeled on the secondary antibody to produce a chemiluminescence reaction, which can sensitively detect the presence of the target protein.ECL substrate chemiluminescence detection kit was developed based on a new generation of enhanced chemiluminescence substrate, and its composition was optimized.Low product background, good stability.It is catalyzed by horseradish peroxidase (HRP) to produce a chemical reaction, which can be exposed to X-ray film, or can be directly detected by Luminometer or fluorescent CCD scanning.

Component

A liquid;B liquid

Protocol

1. According to the conventional Western blot operation, after the incubation of the secondary antibody, during the final washing, according to the size of the membrane, 0.5 mL solution A and 0.5 mL solution B were mixed every 10 cm2 to prepare the working solution for luminescence detection. 2. Remove the film with flat-tipped forceps and touch the lower edge of the film gently with absorbent paper to remove excess liquid on the film.The protein of the film is placed face up on the clean plastic wrap (some plastic wrap on the market may quench the fluorescence when wrapping the imprinted film, so high quality plastic wrap should be selected).Transfer the prepared luminescence detection solution to the protein film with a pipette, cover it evenly, and incubate at room temperature for 2 minutes. 3. Hold the protein film with flat head forceps and touch the lower edge of the film gently with absorbent paper to remove excess liquid from the film.Wrap the egg whites face up in a clean plastic wrap.Gently remove the bubbles and secure them in the X-ray cartridge. 4. Put an X film on the wrapped film in the darkroom, close the cassette, and expose for 30 seconds to 1 minute.Imminently develop and fix the image, shortening or lengthening the exposure time of the next X-ray, depending on its exposure intensity (up to several hours for weak signals).

Operating Instructions

1.ECL liquid A and liquid B must be replaced in the process of suction, the mutual contamination of liquid A and liquid B will lead to gradual failure of liquid A or liquid B, affecting the subsequent use effect. 2. After using each solution, please close the bottle cap tightly to prevent failure.Liquid B, in particular, contains oxidant, which is easy to be reduced and ineffective. 3. The fluorescence of ECL lasts for a long time, but the fluorescence becomes stronger in the first 30 minutes after the reaction, and then the fluorescence gradually decreases. Therefore, please make full use of this 30 minutes of strong fluorescence to press the tablet. 4. Both Liquid A and Liquid B of ECL are harmful to human body. Please pay attention to proper protection during operation. For your safety and health, please wear a lab coat and disposable gloves to operate.

Stability & Storage

Store at 2-8℃ for 6 months away from light.