bchp

Collagen Hybridizing Peptide, Biotin Conjugate (B-CHP)

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Product Description

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.

Applications: immunofluorescence, immunohistochemistry

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).

biotin-special

 

Key Publications

  1. Targeting and mimicking collagens via triple helical peptide assemblies. Curr. Opin. Chem. Biol., 2013. [link]
  2. Targeting collagen strands by photo-triggered triple-helix hybridization. Proc. Natl. Acad. Sci. U.S.A., 2012. [link]
  3. In situ imaging of tissue remodeling with collagen hybridizing peptides. ACS Nano, 2017. [link]
  4. Visualizing collagen proteolysis by peptide hybridization: From 3D cell culture to in vivo imaging. Biomaterials, 2018. [link]
  5. Molecular level detection and localization of mechanical damage in collagen enabled by collagen hybridizing peptides. Nat. Commun., 2017. [link]
  6. Molecular assessment of collagen denaturation in decellularized tissues using the collagen hybridizing peptide. Acta Biomater., 2017. [link]
  7. 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.

 

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Product Citations

  1. Xudong Li et al., Molecular detection and assessment of intervertebral disc degeneration via a collagen hybridizing peptide. ACS Biomaterials Science & Engineering (2019) more info
  2. 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
  3. Stephen Weiss et al., Divergent matrix-remodeling strategies distinguish developmental from neoplastic mammary epithelial cell invasion programs. Developmental Cell (2018) more info
  4. Fiona Watt et al., Fibroblast state switching orchestrates dermal maturation and wound healing. Molecular Systems Biology (2018) more info
  5. Karl Kadler et al., Protection of circadian rhythms by the protein folding chaperone, BiP. bioRxiv (2018) more info
  6. Fiona Watt et al., Loxl2 is dispensable for dermal development, homeostasis and tumour stroma formation. Plos One (2018) more info
  7. 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
  8. Kenneth Monson et al., Detection and characterization of molecular-level collagen damage in overstretched cerebral arteries. Acta Biomaterialia (2018) more info
  9. 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
  10. Samuel Veres et al., In tendons, differing physiological requirements lead to functionally distinct nanostructures. Scientific Reports (2018) more info
  11. 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
  12. Mark Banaszak Holl et al., Fatigue failure mechanism of anterior cruciate ligament fracture. (2018) more info
  13. Themis Kyriakides et al., Decellularized materials derived from TSP2-KO mice promote enhanced neovascularization and integration in diabetic wounds. Biomaterials (2018)
  14. Per Fogelstrand et al., Systematic in vitro comparison of decellularization protocols for blood vessels. Plos One (2018) more info