User Guides | Collagen Hybridizing Peptides | 3Helix, Inc. – 3Helix Inc

Collagen Hybridizing Peptide (CHP) User Guides

Your definitive guides for staining, detecting, and imaging collagen remodeling. Download protocols for specific applications, including histology, automated staining, and small animal imaging. Download protocols for specific applications, including histology, automated staining, and small animal imaging.

Total Collagen
(Histology)

This guide provides a protocol for histological staining of total collagen content using F-CHP, R-CHP, or B-CHP. It requires heat-induced epitope retrieval (HIER) or similar artificial denaturation of all collagens, allowing the CHPs to bind and provide a reliable readout of the total amount.

Damaged Collagen
(Histology)

This guide outlines the histological staining of damaged collagen using F-CHP (green fluorescence), R-CHP (red fluorescence), or B-CHP (biotin conjugate). The key is to avoid heating the tissue sample (no HIER) to ensure the CHPs only bind to the endogenous damaged collagen.

Damaged Collagen
(In Vivo)

This guide focuses on near-infrared fluorescence in vivo imaging of collagen remodeling in small animals using sCy7.5-CHP, which specifically binds to denatured collagen chains in tissues under disease or injury. It includes a non-binding control peptide and reconstitution instructions for injection.

In-Gel Western

This protocol is for the in-gel detection and quantification of collagen content in a biological sample after SDS-PAGE. It uses F-CHP (green) or R-CHP (red) to stain the denatured collagen bands directly in the gel, followed by visualization with a fluorescence gel imager.

Auto-CHP Protocol
(Leica Bond RXM)

This protocol is for automated histological staining of collagen using Auto-CHP (Biotin Conjugate) on platforms like the Leica Bond RXM. It includes methods for staining active remodeling (damaged) collagen (without HIER) and total collagen (with HIER).

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