Collagen Hybridizing Peptide, Biotin Conjugate
Description
The collagen hybridizing peptide (CHP) is a novel and unique peptide that specifically binds unfolded collagen chains, both in vitro and in vivo.[1,2,3] By sharing the Gly-X-Y repeating sequence of natural collagen, CHP has a strong capability to hybridize with denatured collagen chains by reforming the triple helical structure, in a fashion similar to DNA fragments annealing to complementary DNA strands. CHP is extremely specific: it has negligible affinity to intact collagen molecules due to lack of binding sites, and it is inert towards non-specific binding because of its neutral and hydrophilic nature.
CHP is a powerful histopathology tool which enables straightforward detection of inflammation and tissue damage caused by a large variety of diseases, as well as tissue remodeling during development and aging.[3] CHP robustly visualizes the pericellular matrix turnover caused by proteolytic migration of cancer cells within 3D collagen culture, without the use of synthetic fluorogenic matrices or genetically modified cells.[4] CHP can measure and localize mechanical injury to collagenous tissue at the molecular level.[5] It also enables assessment of collagen denaturation in decellularized extracellular matrix.[6] In addition, CHP can be used to specifically visualize collagen bands in SDS-PAGE gels without the need for western blot.[7]
B-CHP is labeled with biotin for avidin/streptavidin-mediated detection. B-CHP offers flexible detection options, such as non-green fluorescence and HRP-enzyme methods, to avoid background and enhance signal.
Specificity: CHP binds to the unfolded triple-helical chains of all collagen types (e.g., I, II, III, IV, etc).[3,7]
Applications: immunofluorescence,[3] immunohistochemistry[3]
Specification
| Synonyms |
B-CHP, collagen mimetic peptide (CMP) |
| Molecular weight |
2762.01 g/mol |
| Purity |
95% by HPLC |
| Conjugate |
Single biotin tag per peptide |
| Content |
Purified lyophilized powder |
| Storage |
-20 °C as powder, 4 °C after reconstitution in water |
Features
- More informative, reliable and convenient than zymography, DQ collagen, SHG, and TEM
- High affinity and unparalleled specificity to collagen with essentially no nonspecific binding
- Applicable to all types of collagen from all species, relying on collagen's secondary structure instead of any defined sequence for binding
- Suitable for both frozen and paraffin-embedded sections with no need for antigen retrieval
- A non-antibody approach with no species restrictions against any co-staining antibody
- Small size (2% of IgG by MW) enabling facile tissue penetration and whole specimen staining without sectioning
- Stable in solution under 4 °C, eliminating the need to aliquot for storage
Special applications
B-CHP provides the flexibility for visualizing damaged collagen using any user-selected fluorophore (e.g., AlexaFluor dyes conjugated streptavidin), or a non-fluorescence method (e.g., through NeutrAvidin-HRP mediated DAB staining).
In addition, B-CHP and commercial streptavidin-conjugated gold nanoparticles can be used in combination to label unfolded collagen molecules under TEM: collagen fibers in an intact or mechanically overloaded rat tail tendon probed by B-CHP and streptavidin conjugated 10nm-sized gold nanoparticles (scale bar: 200 nm).
Key Publications
- Targeting and mimicking collagens via triple helical peptide assemblies. Curr. Opin. Chem. Biol., 2013. [link]
- Targeting collagen strands by photo-triggered triple-helix hybridization. Proc. Natl. Acad. Sci. U.S.A., 2012. [link]
- In situ imaging of tissue remodeling with collagen hybridizing peptides. ACS Nano, 2017. [link]
- Visualizing collagen proteolysis by peptide hybridization: From 3D cell culture to in vivo imaging. Biomaterials, 2018. [link]
- Molecular level detection and localization of mechanical damage in collagen enabled by collagen hybridizing peptides. Nat. Commun., 2017. [link]
- Molecular assessment of collagen denaturation in decellularized tissues using the collagen hybridizing peptide. Acta Biomater., 2017. [link]
- Direct detection of collagenous proteins by fluorescently labeled collagen mimetic peptides. Bioconjug. Chem., 2013. [link]
Additional information
CHP can slowly self-assemble into the triple helical structure in solution during storage. The trimeric CHP requires a simple heating step prior to usage. Please check the protocol for details: CHP User Guide.pdf
For research use only. Not intended or approved for diagnostic or therapeutic use.
Product Citations
- Nikolaos Frangogiannis et al., Protective effects of activated myofibroblasts in the pressure-overloaded myocardium are mediated through Smad-dependent activation of a matrix-preserving program. Circulation Research (2019) more info
- Christopher Fry et al., Anterior cruciate ligament tear promotes skeletal muscle myostatin expression, fibrogenic cell expansion, and a decline in muscle quality. The American Journal of Sports Medicine (2019) more info
- Michele Marino et al., Molecular-level collagen damage explains softening and failure of arterial tissues: A quantitative interpretation of CHP data with a novel elasto-damage model. Journal of the Mechanical Behavior of Biomedical Materials (2019) more info
- Jeffery Molkentin et al., An acute immune response underlies the benefit of cardiac adult stem cell therapy. bioRxiv (2019) more info
- Xudong Li et al., Molecular detection and assessment of intervertebral disc degeneration via a collagen hybridizing peptide. ACS Biomaterials Science & Engineering (2019) more info
- Xudong Li et al., Microfluidic disc-on-a-chip device for mouse intervertebral disc—pitching a next-generation research platform to study disc degeneration. ACS Biomaterials Science & Engineering (2019) more info
- Karl Kadler et al., Protection of circadian rhythms by the protein folding chaperone, BiP. The FASEB Journal (2019) more info
- Stephen Weiss et al., Divergent matrix-remodeling strategies distinguish developmental from neoplastic mammary epithelial cell invasion programs. Developmental Cell (2018) more info
- Fiona Watt et al., Fibroblast state switching orchestrates dermal maturation and wound healing. Molecular Systems Biology (2018) more info
- Fiona Watt et al., Loxl2 is dispensable for dermal development, homeostasis and tumour stroma formation. Plos One (2018) more info
- Matthew Abramowitz et al., Skeletal muscle fibrosis is associated with decreased muscle inflammation and weakness in patients with chronic kidney disease. American Journal of Physiology-Renal Physiology (2018) more info
- Kenneth Monson et al., Detection and characterization of molecular-level collagen damage in overstretched cerebral arteries. Acta Biomaterialia (2018) more info
- Spencer Szczesny et al., Fatigue loading of tendon results in collagen kinking and denaturation but does not change local tissue mechanics. Journal of Biomechanics (2018) more info
- Samuel Veres et al., In tendons, differing physiological requirements lead to functionally distinct nanostructures. Scientific Reports (2018) more info
- Svenja Illien-Junger et al., Dietary advanced glycation end-product consumption leads to mechanical stiffening of murine intervertebral discs. Disease Models & Mechanisms (2018) more info
- Mark Banaszak Holl et al., Fatigue failure mechanism of anterior cruciate ligament fracture. (2018) more info
- Themis Kyriakides et al., Decellularized materials derived from TSP2-KO mice promote enhanced neovascularization and integration in diabetic wounds. Biomaterials (2018)
- Per Fogelstrand et al., Systematic in vitro comparison of decellularization protocols for blood vessels. Plos One (2018) more info
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