Native human collagen type V isolated from human placenta by partial pepsin digestion under acidic conditions, followed by differential salt precipitation. The lyophilized product is =95% type V, with other human collagens at less than 5% combined and non-collagen proteins below 1%. Donor placenta is screened negative for HBsAg, HCV and HIV-1/HIV-2. The native a1(V)2a2(V) heterotrimeric triple helix is preserved through purification — confirmed by SDS-PAGE and by reactivity with type-V-specific anti-collagen antibodies.
Type V is the fibril-regulating collagen of the body's interstitial connective tissue. It co-assembles with type I and type III into the heterotypic fibrils that build skin, bone, tendon, vasculature, liver, lung, muscle and the placental stroma — and it is type V's a-chain stoichiometry, not the absolute amount of collagen, that sets fibril diameter, fibril packing and tissue tensile properties. Loss-of-function mutations in COL5A1 and COL5A2 cause classical Ehlers–Danlos syndrome; abnormal type V expression is a feature of scleroderma, idiopathic pulmonary fibrosis, atherosclerosis, hypertrophic scarring and the corneal-stroma changes of keratoconus. cat. 436 supplies the defined human-source reagent for any cell, biophysical, immunological or autoimmunity experiment where the fibril-regulating collagen is required and a species-matched human standard is needed.
cat. 436 is the human-source counterpart to the bovine type V SKU (cat. 184 atelocollagen, sterile, 3D-gel grade) and the type V partner in the human-collagen autoimmunity antigen panel alongside cat. 288 (type I), cat. 210 (type II), cat. 889 (type III) and cat. 425 (type IV).
SPECIFICATIONS
Catalogue number: 436
Amount: 0.1 mg
Source: Human placenta (HBsAg / HCV / HIV-1 / HIV-2 negative)
Purity: Human collagen type V =95%. Other human collagens <5%. Non-collagen proteins <1%
Format: Lyophilized
Purification: Partial pepsin digestion under acidic conditions; differential salt precipitation
STORAGE / RECONSTITUTION
Dissolve in 0.1 M acetic acid, pH 3.0.
Shipping at ambient temperature. Long-term storage: 2 years at -20 °C or lower.
APPLICATIONS
- Human type V collagen ELISA, immunoblot and dot-blot reference standard — defined human-source antigen for quantifying type V collagen, its a1(V) / a2(V) chains and degradation fragments in serum, urine, synovial fluid, cell-culture supernatants and tissue lysates.
- Antigen for raising anti-human-collagen-type-V antibodies — pure human-source immunogen for rabbit, mouse, goat or chicken immunisation programmes targeting clinical or research-grade anti-type-V antibodies.
- Antigen for anti-collagen-V autoantibody serological assays — quantify pathological IgG, IgM and IgA autoreactivity against human type V collagen in lung-allograft rejection, idiopathic pulmonary fibrosis, scleroderma and post-transplant chronic-rejection diagnostic workups.
- Antigen panel for human-collagen autoimmunity research — paired with cat. 288 (type I), cat. 210 (type II), cat. 889 (type III) and cat. 425 (type IV), cat. 436 completes the human-collagen serology array used in disc-, joint-, dermal-, vascular- and connective-tissue autoimmunity screens.
- Coating substrate for dermal-fibroblast, vascular smooth-muscle-cell, hepatic-stellate-cell and lung-fibroblast culture — type V's fibril-nucleating role in interstitial matrix makes it the appropriate substrate for studying fibrogenic cell phenotypes and matrix-assembly biology in 2D culture.
- Substrate for studies of collagen-I / III / V heterotypic fibril assembly — defined human-source type V for in-vitro reconstitution experiments that probe fibril diameter, fibril packing and a-chain-stoichiometry-controlled mechanical properties.
- Substrate for classical Ehlers–Danlos-syndrome (cEDS / COL5A1 / COL5A2) modelling — defined human substrate for patient-derived fibroblast / iPSC-derived stromal-cell phenotyping and matrix-assembly defect studies.
- Substrate for corneal-stroma and keratoconus research — type V's role in the orderly small-diameter fibril packing of corneal stroma makes it the matched antigen for corneal-collagen quantification, anti-collagen-V autoreactivity screens and keratoconus drug-discovery workflows.
- Substrate for skin-aging, photoaging and dermal-regulation research — defined human standard for quantifying fibrillar-collagen response (types I, III and V together) to nicotinamide, copper, retinoids and other dermal-pharmacology candidates in fibroblast cell models.
- Reference for in-vitro fibrosis and fibroblast-activation assays — defined human substrate for quantifying type V production by activated fibroblasts in scleroderma, pulmonary fibrosis, hepatic fibrosis and tumour-associated stroma research.
- Substrate for type-V-specific protease-activity assays — defined human substrate for MMP-2, MMP-9, MMP-13, cathepsin K and stromal-protease cleavage assays in fibrotic-disease and cancer-invasion drug discovery.
- Reference material for biophysics and structural studies — native a1(V)2a2(V) triple-helical human collagen V for AFM, optical-tweezers, persistence-length, SAXS / SANS and fibril-assembly characterisation work.
APPLICATION HIGHLIGHTS — HIGH-VALUE RESEARCH NICHES
- Human-source antigen for classical Enzyme-Linked Immunosorbent Assay of anti-collagen V autoantibodies in lung-transplant rejection and idiopathic pulmonary fibrosis research.
- Defined substrate for fibrillogenesis studies — collagen V nucleates and limits the diameter of type I collagen fibrils.
- Coating matrix for platelet-adhesion, corneal-stroma and fibroblast-attachment assays.
- Reference standard for quantification of type V collagen in tissue digests, cell lysates and conditioned media.
- Immunogen and QC antigen for anti-human collagen type V antibody development and specificity cross-reactivity panels.
- Model matrix for classical Ehlers-Danlos syndrome (COL5A1 / COL5A2) structure-function research.
REFERENCES
(1) Capossela S., Schläfli P., Bertolo A., Janner T., Stadler B.M., Pötzel T., Baur M., Stoyanov J.V. (2014) — Degenerated human intervertebral discs contain autoantibodies against extracellular matrix proteins. European Cells and Materials 27: 251–263. doi:10.22203/eCM.v027a18.
The authors used cat. 436 as the defined human type V collagen antigen in an ELISA / immunoblot panel for anti-ECM autoantibody screening of serum and disc-tissue extracts from degenerated-disc patients, alongside cat. 288 (human type I) and cat. 889 (human type III). The study demonstrates for the first time the existence of disc-targeted autoimmunity against human collagens including type V — and validates cat. 436 as the species-matched human collagen V antigen for autoimmunity-array work.
(2) Philips N., Chalensouk-Khaosaat J., Gonzalez S. (2015) — Stimulation of the fibrillar collagen and heat-shock proteins by nicotinamide or its derivatives in non-irradiated or UVA-radiated fibroblasts, and direct anti-oxidant activity of nicotinamide derivatives. Cosmetics 2(2): 146–161. doi:10.3390/cosmetics2020146.
The authors used cat. 436 as the human type V collagen reference standard for dot-blot / ELISA quantification of fibrillar-collagen response (types I, III and V together) in dermal fibroblasts treated with nicotinamide and its derivatives, with and without UVA radiation. The work positions cat. 436 as the human-source type V quantification standard for cosmeceutical and anti-photoaging fibroblast-pharmacology work.
(3) Philips N., Samuel P., Parakandi H., Gopal S., Siomyk H., Ministro A., Thompson T., Borkow G. (2012) — Beneficial regulation of fibrillar collagens, heat-shock protein-47, elastin fibre components, transforming growth factor-ß1, vascular endothelial growth factor and oxidative stress effects by copper in dermal fibroblasts. Connective Tissue Research, 1–7.
The authors used cat. 436 as the human type V collagen ELISA / dot-blot standard in a study of copper-driven regulation of fibrillar collagens, HSP-47, elastin, TGF-ß1 and VEGF in dermal fibroblasts. Establishes cat. 436 alongside cat. 288 (type I) and cat. 889 (type III) as the standard human-collagen quantification reference panel for fibroblast-pharmacology screens in dermal-aging and wound-healing research.
