What is ECL Western blotting Substrate?

The ECL Western Blotting Substrate is a highly sensitive non-radioactive, enhanced luminol-based chemiluminescent substrate for the detection of horseradish peroxidase (HRP) on immunoblots.

What substrate is used in Western BLoT?

The Western BLoT Quant HRP Substrate is a horseradish peroxidase (HRP) substrate specially developed for CCD imaging of Western blots. The signal from Western BLoT Quant is linear with respect to protein amount over a broad range of concentrations, allowing full use of the linear range of CCD detection.

What is chemiluminescent substrate?

Chemiluminescent substrates for HRP are most commonly two-component systems consisting of a stable peroxide solution and an enhanced substrate solution, often luminol-based. In most cases, to make a working solution, equal volumes of the two components are mixed together.

How does ECL HRP work?

ECL Uses an HRP Substrate for High-Sensitivity Western Blot Detection. Chemiluminescence assays are commonly used for the detection of proteins on western blots. The HRP substrate luminol emits light at 428 nm; however, luminol emits light only weakly, so enhancers are added to the reaction to increase the signal.

What is HRP substrate?

Horseradish peroxidase (HRP) substrates are used in the immunostaining and immunodetection of specific proteins and antigens. HRP is a widely used label, often conjugated to secondary antibodies in immunochemical applications such as immunohistochemistry, western blotting, and ELISA.

Is ECL toxic?

Inhalation: May be harmful if inhaled. May cause respiratory tract irritation. Skin: May be harmful if absorbed through skin.

What is PVDF membrane?

PVDF (polyvinylidene difluoride) membranes are widely used for immunoblotting techniques such as Western blots. During Western blots, samples are transferred from a protein gel onto the PVDF membrane by electroblotting, and then used for immunodetection or protein sequencing.

What is ECL reagent?

Amersham™ ECL™ Western Blotting Detection Reagents were the world′s first commercial chemiluminescent reagents for Western blotting and are still the most widely used. They are at least 10-fold more sensitive than colorimetric assays and produce excellent and reliable results every time.

What is western blot chemiluminescence?

Chemiluminescent western blot detection is a highly sensitive alternative to isotopic detection. Instead of radioactively labeled antibodies, enzyme-conjugated antibodies are used to convert a substrate to one that produces a light signal. The signal can be captured on film or by dedicated imaging equipment.

What is the specific substrate for HRP?

Colorimetric substrates for ELISA applications

Substrate Enzyme Absorbance and color
OPD Substrate HRP 490 nm (450 nm) Green (Orange)
PNPP (p-Nitrophenyl Phosphate)
1-Step PNPP Substrate Solution AP 405 nm Yellow
PNPP Substrate AP 405 nm Yellow

What is the substrate for horseradish peroxidase?

The enzyme horseradish peroxidase (HRP), found in the roots of horseradish, is used extensively in biochemistry applications. It is a metalloenzyme with many isoforms, of which the most studied type is C. It catalyzes the oxidation of various organic substrates by hydrogen peroxide….Horseradish peroxidase.

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What is HRP in Western blot?

Enzymatic labels are most commonly used for Western blotting and, although they require extra steps, can be extremely sensitive when optimized with an appropriate substrate. Horseradish peroxidase (HRP) is the enzymes used most extensively as labels for protein detection.