ECLiNano™ ECL Chemiluminescence Kit

ECL03-01

Description

ECLiNano™ ECL Chemiluminescence Kit is a sensitive chemiluminescence kit based on luminol ECL chemiluminescence. This product can react with horseradish peroxidase (HRP) conjugated on secondary antibody to produce fluorescence. Samples are examined by X-ray lithography or other appropriate fluorescence imaging equipment (CCD camera, etc.). It is used for  developing Western blots with high sensitivity and more stable color development, which was suitable for the detection of mid to mid to high abundance target protein. This product is indicated for the detection of target proteins is less than or equal to nanogram range.

Best used together with our ECLiPico™ (picogram) and ECLiFemto™ (femtogram) high sensitivity and very high sensitivity ECL substrates to titrate variable sensitivity needs for your Western blot analyses. 

Storage and Handling Conditions

Transport with wet ice; Store at 4℃ away from light, valid for 12 months. If not used for a long time, it can be stored at -20℃.

Component

Kit Quantity	ECL03 ECL A solution	ECL03 ECL B solution
100mL	50 mL	50 mL
200mL	100 mL	100 mL
800mL	400 mL	400 mL

Assay Protocol

1. Preparation of hypersensitive ECL working solution: Mix the ECL A solution and ECL B solution in equal volumes, and store at 4℃away from light. This mixture should be prepared before use.
2. In Western experiment, PVDF membrane was incubated with secondary antibody, washed several times, and excess liquid was 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. Adjust exposure conditions according to luminous intensity.

Note:

1. High sensitive ECL A liquid and high sensitive ECL B liquid in the pipetting process, the pipette tips must be replaced. The reciprocal 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 pipette tip. 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, it may be that the target protein expression is very weak, which can prolong the tablet pressing time.

5. Wear a lab coat and disposable gloves during operation.