Westerns Done Right with Q-Bright
Q-Bright Chemiluminescent Detection Kits - Which Q-Bright Is Best For You?
Q-Bright chemiluminescent detection reagent is a luminol-based substrate for detection of horseradish peroxidase (HRP) conjugates in western blot applications. Multiple products and sizes allow you to choose the right product for your needs.
Robust Signal for Routine Western Blotting
Comparable Sensitivity to
SuperSignal™ West Pico
The Q-Bright line of substrates is designed to provide options for protein detection across a range of sensitivities. Here, we demonstrate the differing sensitivities with identical primary and secondary antibody concentrations and imaging time. The western blotting conditions can be further optimized for maximal signal and signal to noise by adjusting the antibody concentrations and exposure times. GAPDH His-Tag Protein Standard was loaded at 100ng-100pg per well and transferred to a PVDF membrane. The membrane was incubated with Rabbit anti-GAPDH polyclonal antibody (1:1000) followed by Goat anti-Rabbit Horseradish Peroxidase (HRP) secondary antibody with minimal cross-reactivity to murine, human, bovine and horse IgGs (11-0201-0503 1:50,000). The membranes were incubated with Q-Bright Enhance Plus, Endure, Endure Plus or Extreme for 5 minutes then imaged on a digital chemiluminescent imaging system with a 60 second exposure time.
Introducing Enhance Plus, the newest product to the Q-Bright line of Chemiluminescent Substrates
Q-Bright Enhance Plus is more sensitive than many of the standard, commercial ECL reagents and is useful for routine western blots designed to detect abundant proteins.
The Q-Bright Enhance Plus Chemiluminescent Detection Kit provides a strong signal for abundant proteins. Here, we demonstrate the differing sensitivities with identical primary and secondary antibody concentrations and imaging time. The western blotting conditions can be further optimized for maximal signal and signal to noise by adjusting the antibody concentrations and exposure times. HeLa cell lysate was loaded at 10-1.21ug/well and transferred to a PVDF membrane. The membrane was incubated with Rabbit anti-Beta-Catenin (1:10,000) followed by Goat anti-Rabbit Horseradish Peroxidase (HRP) secondary antibody with minimal cross-reactivity to murine, human, bovine and horse IgGs (11-0201-0503 1:20,000). Finally, the membranes were incubated with Q-Bright Enhance Plus or a commercial ECL reagent for 5 minutes then imaged together on a digital chemiluminescent imaging system.
Q-Bright Endure provides a long lasting, stable signal over hours allowing for repeat imaging of the same blot and longer exposure times.
Cell lysate was loaded at 500-15ng total protein/well and transferred to a PVDF membrane. The membrane was incubated with Rabbit anti-b Catenin primary antibody (1:10,000) followed by Goat anti-Rabbit IgG (H&L), Horseradish Peroxidase (HRP) min x mouse, human, bovine, horse IgG (11-0201-0503; 1:25,000). To develop the signal, the membrane was incubated in Q-Bright Endure working solution for 5 minutes before transferring to the digital imager. The membrane was imaged with a 1 minute exposure time at baseline (0h), followed by repeat images with identical exposure settings at 1h, 2h, 3h, and 5h to demonstrate the sustained signal provided by Q-Bright Endure.
Q-Bright Endure Plus and Q-Bright Extreme provide the highest levels of sensitivity
Use the most sensitive Q-Bright chemiluminescent reagents to detect low abundant proteins or smaller quantities of starting material. Q-Bright Endure Plus provides great sensitivity without extensive optimization of western blotting conditions that is required when working with the most sensitive of reagents. The sensitive reagents conserve resources by using less primary and secondary antibody.
Optimal Signal and Dynamic Range in Chemiluminescent Western Blot
Primary and secondary antibody concentrations can be adjusted to provide an optimal signal and dynamic range for the end user.
Primary Antibody Dilution Range
Secondary Antibody Dilution Range
Cell Lysates 1.5-0.19 ug/well
PVDF Membrane
Primary: Mouse anti-b actin 1:5,000, 1:25,000
Secondary: Goat anti-Mouse HRP (11-0101-0303) 1:50,000
Q-Bright: Endure Plus
Exposure: 15 seconds
Cell Lysates 1.5-0.19 ug/well
PVDF Membrane
Primary: Mouse anti-b actin 1:10,000
Secondary: Goat anti-Mouse HRP (11-0101-0303) 1:50,000, 1:100,000 and 1:200,000
Q-Bright: Endure Plus
Exposure: 15 seconds
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