- The collagen hybridizing peptide (CHP) is a novel and unique peptide that specifically binds unfolded collagen chains, bothin vitroand 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.CHP robustly visualizes the total collagen content of tissue sections that have undergone heat-induced epitope retrieval (HIER). The ratio between damaged collagen and total collagen can be a useful metric for evaluating disease/damage progression over time. Additionally, the signal can be quantified using image analysis software including ImageJ for fluorescent quantification (See Application Note)
- The Auto B-CHP incorporates our newest sequence which allows them to be used with automated staining platforms that are “Open” such as the Bond-RXm (Leica), Discovery Ultra (Roche), and Valent (Biocare Medical) systems. The new sequence will not refold over the course of tissue staining and they do not require a pre-activation heating step. The Auto B-CHP is a powerful histology 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.  The compatibility with numerous “Open” automated staining platforms enables easy, consistent, and reproducible staining that can be done simultaneously with other stains.
- Auto B-CHP is labeled with biotin for avidin/streptavidin-mediated detection. The biotin label offers flexible detection options, such as non-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]
|Synonyms||Auto B-CHP, Auto Biotin-CHP, collagen mimetic peptide (CMP)|
|Molecular Weight||2981.19 g/mol|
|Purity||95% by HPLC|
|Conjugate||Single biotin tag per peptide|
|Content||Purified lyophilized powder, white color|
|Storage||-20 °C as powder (long term storage), 4 °C after reconstitution in water|
Unheated Tissue- Active Remodeling Collagen
Heated Tissue- Total Collagen
Fibrotic mouse liver sections (FFPE) stained with Auto-CHP. These sections were taken from mice induced with the carbon tetrachloride (CCl4) fibrosis model. Sections were deparaffinized following a standard protocol and stained on an automated Leica Bond-RXm platform using the Auto-CHP which contains a biotin label. The damaged or remodeling collagen is stained brown with the use of a streptavidin- diaminobenzidine (DAB). The unheated section (left) shows the active remodeling (damaged) collagen present in the tissue due to the progression of fibrosis; dark brown areas indicate regions with high collagen turnover. The heated section (right) had all the collagen in the sample purposefully denatured prior to staining with Auto-CHP. This represents the total amount of collagen present in the serial section. The ratio between total and damaged collagen may offer insight into disease progression.
- New sequence designed for use on automated platforms does not require any pre-activation (pre-heating) step unlike CHPs for manual histology
- Biotin label offers flexibility for imaging numerous fluorescent wavelengths or non-fluorescent imaging with HRP for signal amplification
- Can be used on “Open” systems such as the Bond-RX or RXm (Leica), Valent or IntelliPATH FLX (Biocare Medical), Discovery Ultra (Roche), Autostainer 480S (ThermoFisher), or AS Link 48 (Dako/Agilent)
- High affinity and unparalleled specificity to collagen turnover 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
- Stable in solution under 4 °C for months, eliminating the need to aliquot for storage