Collagen type III is a fibrillar collagen of the major vascular and visceral extracellular matrix — the dominant structural collagen of large blood vessels, the uterus, the bowel and reticular networks throughout dermis, lung and lymphoid tissue. Heterozygous COL3A1 mutations cause vascular Ehlers-Danlos syndrome, and type III is the obligate co-fibrillating partner of type I in nearly every soft-tissue heterotypic I/III fibril. The accurate, species-matched human form of the protein is essential whenever the experimental readout depends on the human ECM signature: human-cell ECM microarrays, primary human-cell substrate work, human-fibroblast scar / fibrosis modelling, anti-human-collagen autoantibody quantification, and human-tissue WB / dot-blot reference loading.
YO Proteins cat. 889 is native human collagen type III purified from human placenta. The purification chain combines partial pepsin digestion in acidic conditions, differential salt precipitation, and DEAE ion-exchange chromatography, delivering = 90 % type III with type I < 10 % and all other collagen types and non-collagen contaminants below 1 %. Donor placental tissue is screened HBsAg / HCV / HIV-1 / HIV-2 negative; the preparation is supplied as a salt-free lyophilised powder for reconstitution in 0.5 M acetic acid pH 2.5.
SPECIFICATIONS
Catalogue number: 889
Product name: Collagen type III (human)
Source: Human placenta. Donor pool tested negative for HBsAg, HCV, HIV-1 and HIV-2
Purity: Human collagen type III = 90 %; human collagen type I < 10 %; other human collagen types and non-collagen proteins < 1 %
Format: Lyophilised, salt-free
Purification: Partial pepsin digestion in acidic conditions ? differential salt precipitation ? DEAE chromatography
Gene symbol: COL3A1 (chromosome 2q32)
UniProt: P02461
Package size: 0.1 mg
Grade: Research use only (not for diagnostic or therapeutic use)
STORAGE / RECONSTITUTION
Reconstitute in 0.5 M acetic acid, pH 2.5. Mix gently overnight at +4 °C — a low-speed orbital shaker or end-over-end rotator is ideal; do not vortex. A homogeniser may be required to disperse the recalcitrant sediment, which is normal behaviour for high-purity native fibrillar collagen and does not indicate aggregation damage.
The acid-reconstituted material retains the immunological reactivity of native collagen type III and is competent for in-vitro fibrillogenesis after neutralisation under appropriate conditions (controlled raise to pH 7.4, isotonic salt). For ELISA coating, perform a 1:100 dilution into PBS or carbonate / bicarbonate coating buffer (pH 9.6) immediately before pipetting onto the plate. For Western-blot positive-control loading, use 50–200 ng per lane under reducing conditions. For nitrocellulose-slide microarray printing, dilute the acid stock into the printing buffer specified by the slide vendor.
For cell-culture use, sterile-filter the diluted working solution (0.22 µm) before contact with cells.
Storage
Shipped at ambient temperature.
Lyophilised protein: store up to 2 years at -20 °C or lower.
Reconstituted protein (0.5 M acetic acid pH 2.5): store at -20 °C in single-use aliquots; avoid repeated freeze–thaw cycles.
APPLICATIONS
- Collagen type III standard for cell-based assays, ELISA, Western blot and dot blot — used as the absolute-quantification reference in 5+ published studies
- Antigen for the production and titration of anti-human collagen III monoclonal and polyclonal antibodies (used in the generation of YO cat. 153 / cat. 380 / cat. 661 anti-collagen-III antibodies)
- Coating material for cell-culture studies — primary human fibroblasts, hepatic progenitors, lung-cancer cells, peripheral nerve / Schwann-cell models
- Coating substrate for nitrocellulose film slides and conjugated-ECM polyacrylamide (ECM-PA) microarrays used for ECM-context cell phenotype mapping
- Coating substrate for microfluidic cell-migration devices investigating ECM-dependent migration speed, directional bias and matrix-degradation phenotypes
- ECM substrate for combinatorial microenvironmental screens of stem-cell / progenitor-cell differentiation (hepatic, mesenchymal)
- ECM substrate in lung-tumour-cell drug-response cell microarrays for ECM-context pharmacology
- Substrate for THP-1 monocyte / M1-macrophage polarisation assays
- Detection antigen for anti-collagen autoantibody quantification (intervertebral-disc autoimmunity, autoimmune connective-tissue disorders)
- Reference protein for advanced-glycation-end-product (AGE) modification studies and AGE-derived artefact characterisation in immunoenzymatic methods
- Reference protein for matrix-metalloproteinase / cardiac wound-healing studies
- Reference protein for peripheral-myelin / Schwann-cell ECM studies
- Reference protein for organ-development ECM mapping (bladder detrusor, vascular adventitia, uterine)
- Component for in-vitro reconstitution of human-matched heterotypic I/III collagen fibrils (combine with YO cat. 288 human collagen I)
APPLICATION HIGHLIGHTS — HIGH-VALUE RESEARCH NICHES
- Species-matched human ECM — native placental collagen III for human-cell substrate work where the bovine form (cat. 169) would confound the readout.
- Vascular Ehlers-Danlos and COL3A1 research — reference antigen for autoantibody and structural studies.
- Heterotypic fibril reconstitution — in-vitro assembly of human I/III fibrils with collagen I cat. 288.
- ECM microarray and microfluidic migration platforms — defined coating substrate for phenotype and motility mapping.
- Antibody development and QC — immunogen and coating antigen for the anti-collagen-III panel cat. 153, 380 and 661.
- Fibrosis and scar modelling — quantitative type III reference in human fibroblast and hepatic-progenitor systems.
REFERENCES
The following peer-reviewed publications use YO Proteins human collagen type III (cat. 889) as the antigen / coating substrate / standard of record.
1. Wegener H, Leineweber S, Seeger K. The vWFA2 domain of type VII collagen is responsible for collagen binding. Biochemical and Biophysical Research Communications. 2012 Dec 10. PMID 23237810.
Used cat. 889 as the binding partner in von Willebrand factor type A (vWFA2) domain affinity assays — mapping the molecular basis of type-VII anchoring-fibril binding to interstitial collagen III.
2. Capossela S, Schläfli P, Bertolo A, Janner T, Stadler BM, Pätzel T, Baur M, Stoyanov JV. Degenerated human intervertebral discs contain autoantibodies against extracellular matrix proteins. European Cells and Materials. 2014 Apr 4;27:251–263. PMID 24706108.
Used cat. 889 alongside YO cat. 288 (collagen I), cat. 210 (collagen II), cat. 425 (collagen IV) and cat. 436 (collagen V) as the immobilised antigen panel for the detection of IgG autoantibodies against human ECM proteins in degenerated intervertebral discs. The defining paper for the YO human-placental-collagen autoantibody panel; the panel is sold together as a bundle for connective-tissue / autoimmunity laboratories.
3. Philips N, Chalensouk-Khaosaat J, Gonzalez S. Stimulation of the fibrillar collagen and heat-shock proteins by nicotinamide or its derivatives in non-irradiated or UVA-irradiated fibroblasts, and direct anti-oxidant activity of nicotinamide derivatives. Cosmetics. 2015;2(2):146–161. doi:10.3390/cosmetics2020146.
Used cat. 889 (with cat. 288 collagen I) as the Western-blot quantification standard for fibrillar collagen induction by nicotinamide derivatives in human dermal fibroblasts under UVA stress. Cosmeceutical / skin-fibroblast application.
4. Kuzan A, Chwilkowska A, Maksymowicz K, Szydelko-Bronowicka A, Stach K, Pezowicz C, Gamian A. Advanced glycation end products as a source of artifacts in immunoenzymatic methods. Glycoconjugate Journal. 2018;35(1). doi:10.1007/s10719-017-9805-4. PMID 29305778.
Used cat. 889 to characterise AGE-derived ELISA artefacts on collagen-type-III-coated plates — methodologically important reference for any lab running anti-AGE / anti-collagen-AGE immunoassays.
5. Pal S, Konkimalla VB. Data on sulforaphane treatment mediated suppression of autoreactive, inflammatory M1 macrophages. Data in Brief. 2016. doi:10.1016/j.dib.2016.03.105. PMID 27222853.
Used cat. 889 as one of the ECM-coating substrates that polarised THP-1 monocyte differentiation toward an M1 inflammatory macrophage phenotype, providing the matrix context for sulforaphane immunomodulation testing.
6. Kaylan KB, Ermilova V, Yada RC, Underhill GH. Combinatorial microenvironmental regulation of liver progenitor differentiation by Notch ligands, TGF-ß, and extracellular matrix. Scientific Reports. 2016;6:23490. PMID 27025873.
Cat. 889 was one of the ECM components in the printed-microarray screen mapping liver-progenitor lineage choice as a function of immobilised ECM identity. Methodology-defining paper for ECM-microarray progenitor-differentiation work.
7. Kourouklis AP, Kaylan KB, Underhill GH. Substrate stiffness and matrix composition coordinately control the differentiation of liver progenitor cells. Biomaterials. 2016;99:82–94. PMID 27235994.
Cat. 889 used in a polyacrylamide-stiffness-modulated ECM-conjugation array to dissect the stiffness × ECM-composition interaction in liver-progenitor differentiation.
8. Kaylan KB, Gentile SD, Milling LE, Bhinge KN, Kosari F, Underhill GH. Mapping lung tumor cell drug responses as a function of matrix context and genotype using cell microarrays. Integrative Biology. 2016;Issue 12. PMID 27796394.
Cat. 889 used in printed ECM-microarrays to map lung adenocarcinoma drug responses as a function of matrix-context — translational pharmacology platform.
9. Meschiari CA, Jung M, Iyer RP, Yabluchanskiy A, Toba H, Garrett MR, Lindsey ML. Macrophage overexpression of matrix metalloproteinase-9 in aged mice improves diastolic physiology and cardiac wound healing after myocardial infarction. American Journal of Physiology. 2018. doi:10.1152/ajpheart.00453.2017. PMID 29030341.
Cat. 889 used as the WB / dot-blot standard for collagen type III quantification in aged-mouse cardiac MMP-9-overexpression infarct-wound-healing studies.
10. Ackerman SD, Luo R, Poitelon Y, Mogha A, Harty BL, D'Rozario M, Sanchez NE, Lakkaraju AKK, Gamble P, Li J, Qu J, MacEwan MR, Ray WZ, Aguzzi A, Feltri ML, Piao X, Monk KR. GPR56/ADGRG1 regulates development and maintenance of peripheral myelin. Journal of Experimental Medicine. 2018 Mar;215(3):941–961. DOI 10.1084/jem.20161714. PMID 29367382.
Cat. 889 used in the analysis of peripheral myelin ECM and the role of the adhesion-GPCR GPR56/ADGRG1 in Schwann-cell biology.
11. Smeulders N, Woolf AS, Wilcox DT. Extracellular matrix protein expression during mouse detrusor development. Journal of Pediatric Surgery. 2003;38(1). PMID 12592609.
Cat. 889 used as the human collagen type III WB standard in a developmental study of bladder-detrusor ECM expression.
