Application:
Substrate of Horseradish peroxidase
for chemiluminescent detection of protein in Western Blot
Features:
▪ Chemiluminescence lasts for up to 6 hours
without significant attenuation
▪ Extremely sensitive able to detect 0.1 pg
protein with very low background
▪ Extremely stable for storage up to 12 mouths at
4 °C
▪ Working solution stable up to 72 hours at
4 °C
Comparisons of
LumiGOLD™ ECL Western Blotting Detection Kit with Other Leading Brand Products:
A.
LumiGOLD™ ECL Western
Blotting Detection of EGFP (enhanced green fluorescent protein) in Wide Linear
Range*
*
10 two-folder serial dilutions of purified EGFP (0.1 pg~51.2 pg in lane 1~10
from left to right, upper panel). EGFP was separated by SDS-PAGE electrophoresis
followed by transfer to a PVDF membrane. The membrane was blocked with 5% skim
milk (product # SL100317) and washed with TBST followed by incubation of mouse
anti-EGFP monoclonal antibody. HRP-conjugated goat anti-mouse IgG was used to
recognize the mouse anti-GFP. The blot was exposed to Fuji film for 20 seconds.
The linear range of LumiGOLDD™ ECL kit was determined (lower panel) by addition
of 10 two-folder serial diluted horseradish peroxidase (HRP) to the LumiGOLD™
working solution followed by determination of luminosity by Beckman Coulter
LD400 luminometer
B. A comparison of LumiGOLD™ ECL
Western Blotting Detection Kit with Popular Brand Products
Products |
Noise* |
Signal** |
Signal / Noise |
LumiGOLD™ |
18905 |
1938181 |
103 |
Amersham ECL™ |
1495 |
93098 |
62 |
Amersham ECL™ Plus |
320478 |
3480213 |
10.9 |
Pierce Pico Femto™ |
362103 |
3353736 |
9.3 |
*
Noise refers to the background chemiluminescence of the working solution without
addition of horseradish peroxidase (HRP)
** Signal refers to the HRP catalyzed chemiluminescence of the working solution after addition of 5 µl
x HRP-conjugated goat anti-mouse IgG (100,000
dilution). Luminosity was determined by a Beckman Coulter LD400 luminometer
C. LumiGOLD™ ECL Western Blotting Detection of
beta-Actin in Comparison of Amersham ECL Kit*
Amersham ECL™
LumiGOLD™
* HEK293
cell lysate was prepared with CompLysis™ Protein Extraction Reagent (Cat #
SL100319) and the same amount of total proteins were loaded to the corresponding
wells (lane #1=lane #5 / lane #2=lane #6 / lane #3=lane #7 / lane #4=lane #8).
beta-Actin was picked up by anti-actin antibody
followed by development with ECL reagents for 2 minutes. The exposure time was
labeled in the above images
Data Sheet
Testimonials
I have used it for several times, it is good according to the results. It seems
that this product is better than others to some degree...
------by Dr. Aibing Wang, NHLBI, NIH