VisiGlo™ HRP Chemiluminescent Substrate Kits

Solutions：Proteomics,Immunology,Molecular Biology,Cell Biology

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AMRESCO’s VisiGlo™ Select HRP Chemiluminescent Substrate Kit combines a broad linear dynamic range and attomolar sensitivy to enable quantitation of low-abundance targets labeled with horseradish peroxidase (HRP). The high sensitivity and signal-to-noise ratio allow for target detection with minimal consumption of valuable protein and antibody reagents, thereby lowering experimental costs. Low-intensity signals undetectable with competing chemiluminescent substrates may be revealed as well, because background levels remain low even for lengthy exposure times . The signal generated by substrate incubation is stable for hours and may be captured by conventional x-ray film, CCD and fluorescence-based imaging systems.

·Highest sensitivity - ideal for detecting low-abundance protein

·Quantitative with broad dynamic range

·High signal-to-noise ratio

·Signal lasts hours - enables repeated exposures and imaging delays

·Versatile imaging with CCD-, x-ray film, and fluorescence-based imagers

Sensitive HRP detection with VisiGlo™ Select HRP Chemiluminescent Substrate. Identical slot-blots with 10-fold serial dilutions of horseradish peroxidase (10 ng - 10 fg) were incubated separately in three chemiluminescent substrates for comparison; VisiGlo™ Select HRP Chemiluminescent Substrate Kit (1B1583, AMRESCO). Amersham™ ECL™ Select (GE) and SuperSignal® West F

emto (Pierce). VisiGlo™ Select sensitivity was comparable to that of competing substrates, with detection of as little as 10 fg HRP.

VisiGlo™ Select provides a long-lasting chemiluminescent signal.

A blot containing HRP bands was incubated 5 minutes with VisiGlo™ Select HRP Chemiluminescent Substrate and imaged at 0, 1 and 3 hours post-

substrate incubation using the same exposure settings. Significant chemiluminescent signal was retained even after 3 hours.

AMRESCO’s VisiGlo™ Prime HRP Chemiluminescent Substrate Kit delivers a wide dynamic range of HRP (horse radish peroxidase) detection that is linear over three orders of magnitude, enabling accurate and quantitative comparison of proteins, especially when combined with CCD imaging. Highly sensitive VisiGlo™ Prime is ideal for detection of low abundance proteins, with sensitivity down to attomolar levels. High abundance proteins are detected without exhibiting substrate depletion, an important feature for simultaneous detection of low and high abundance proteins in a single exposure. The chemiluminescent emission from VisiGlo™ Prime is highly stable, lasting hours after substrate incubation to allow time for multiple exposures to obtain optimal blotting images.

·Attomolar sensitivity

·Quantitative - linear range of signal spans 3 orders of magnitude

·Low background for optimal signal intensity

·Sustained signal allows detection hours after substrate incubation

·Compatible with CCD- and x-ray film based imaging systems

Sensitive HRP detection with VisiGlo™ Prime HRP Chemiluminescent Substrate. Identical slot-blots with 10-fold serial dilutions of horseradish peroxidase (100 ng - 100 fg) were incubated separately in three chemiluminescent substrates for comparison; VisiGlo™ Prime HRP Chemiluminescent Substrate Kit (1B1581, AMRESCO). Amersham™ ECL™ Prime (GE) and SuperSignal® West Dura (Pierce). VisiGlo™ Prime was the only substrate not to exhibit inverse staining of the 100 ng HRP band, a phenomenon associated with substrate burn-out. VisiGlo™ Prime also had greater sensitivity, with a lower limit of HRP detection at 100 fg.

High sensitivity and low background on a Western blot detected with VisiGlo™ Prime. Serial dilutions of K562 cytoplasmic lysate were resolved by SDS-PAGE and transferred to PVDF using Rapid Transfer Buffer (N789). The Western blot was probed with anti-NF-KB (1:2,000) primary and HRP-labeled secondary antibodies. VisiGlo™ Prime HRP Chemiluminescent Substrate (1B1581) was used to detect the target, with NF-KB detected in as little as 625 ng of lysate.

VisiGlo™ Prime provides a long-lasting chemiluminescent signal. A blot containing HRP bands was incubated 5 minutes with VisiGlo™ Prime HRP Chemiluminescent Substrate (1B1581) and imaged at 0, 1 and 3 hours post-substrate incubation using the same exposure settings. Significant chemiluminescent signal was retained even after 3 hours.

AMRESCO's VisiGlo™ HRP Chemiluminescent Substrate Kit is a luminol-based substrate designed for the rapid and sensitive detection of peroxidase-labeled conjugates. It offers increased sensitivity compared to chromogenic substrates in both blotting and ELISA applications. VisiGlo™ can be detected using x-ray film and chemiluminescent imagers in blotting applications or using a luminometer in ELISA. Supplied as kit with equivalent volumes Substrate A and Substrate B for the detection of ~2400 cm2 of membrane. Mix directly before use and apply to blot.

·Luminol-based substrate for rapid detection of peroxidase-labeled conjugates

·Highly sensitive substrate saves precious protein samples and antibodies

·Low background without compromising signal intensity

Detection of ovalbumin with VisiGlo™ HRP Chemiluminescent Substrate Kit. Two-fold serial dilutions of ovalbumin (960 ng - 20 ng) were separated on a 12% Fluorescent SPRINT NEXT GEL® (M318) and then transferred and blotted with ovalbumin antibody using a Rapid Western Blotting Kit (N790). The blot was developed with VisiGlo™ HRP Chemiluminescent Substrate Kit (N218) and the image captured with a Syngene gel documentation system.