3Helix CHP Application: Collagen Identification

SDS-PAGE: Detecting collagen in gels

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Introduction

3Helix Collagen Hybridizing Peptides (CHPs) are short peptides that selectively target damaged and remodeling collagen for applications in therapeutics, cosmetics, drug delivery systems, and life science tools.

Summary

Collagen Identification Application: CHPs enable facile, sensitive, and specific collagen detection in SDS-PAGE gels by its triple-helical hybridization with SDS/heat-denatured collagen during electrophoresis. This simple method avoids Western blot steps (e.g., membrane transfer, blocking), saving time, effort, and materials.

Application Note

Fluorescent F-CHP reliably measures a dilution series of type I collagen directly in an SDS-PAGE gel, and detects a band containing as little as 5 ng of protein (left). CHP identifies multiple collagen types (middle) and has no affinity to non-collagenous molecules in the ECM (middle) and cell lysate (right) due to its neutral and hydrophilic nature. Fluorescence images recorded with a Typhoon Imager (λex: 488 nm). Images were adapted from ref [1] with permission.

Fluorescent F-CHP reliably measures a dilution series of type I collagen directly in an SDS-PAGE gel, and detects a band containing as little as 5 ng of protein (left). CHP identifies multiple collagen types (middle) and has no affinity to non-collagenous molecules in the ECM (middle) and cell lysate (right) due to its neutral and hydrophilic nature. Fluorescence images recorded with a Typhoon Imager (λex: 488 nm). Images were adapted from ref [1] with permission.
In addition, CHP can be used in replacement of unsuccessful collagen antibodies or conventional collagen stains to detect collagen in highly denatured tissues.[1] Images below show complete collagen content in a thoroughly heated porcine ligament section visualized by F-CHP or B-CHP (further detected by HRP-conjugated NeutrAvidin). CHP stain offers higher specificity to collagens than conventional collagen stains such as Sirus Red, which relies on positive charges for collagen binding and can stain other proteins with high content of basic amino acids nonspecifically.[2]
In addition, CHP can be used in replacement of unsuccessful collagen antibodies or conventional collagen stains to detect collagen in highly denatured tissues.[1] Images below show complete collagen content in a thoroughly heated porcine ligament section visualized by F-CHP or B-CHP (further detected by HRP-conjugated NeutrAvidin). CHP stain offers higher specificity to collagens than conventional collagen stains such as Sirus Red, which relies on positive charges for collagen binding and can stain other proteins with high content of basic amino acids nonspecifically.[2]

Reference

  1. Direct detection of collagenous proteins by fluorescently labeled collagen mimetic peptides. Bioconjugate Chemistry (2013).
  2. Sirius red and acid fuchsin staining mechanisms. Biotechnic & Histochemistry (1998) 73, 71–77.

Ilex Life Sciences LLC is an authorized distributor of CHP products manufactured by 3Helix, Inc.