Native human collagen type IV isolated from human placenta by partial pepsin digestion under acidic conditions, followed by differential salt precipitation. The lyophilized, salt-free product is 95% type IV, with other human collagens at less than 5% combined and non-collagen proteins below 0.5%. Donor placenta is screened negative for HBsAg, HCV and HIV-1/HIV-2.
Type IV is the network-forming collagen of basement membranes — the thin specialised ECM that underlies every epithelium and endothelium, surrounds skeletal-muscle fibres and Schwann cells, and forms the glomerular basement membrane (GBM) of the kidney filter. It is the binding ligand for integrins a1ß1, a2ß1, a3ß1 and aVß3, the autoantigen in Goodpasture syndrome (anti-NC1 antibodies against the a3 chain), and a target of mutation in Alport syndrome (COL4A3, COL4A4, COL4A5). cat. 425 supplies the human-source reagent for any cell, biophysical, immunological or matrix-assembly experiment where the defining basement-membrane collagen is required and a species-matched human standard is needed to avoid cross-species epitope mis-detection.
cat. 425 is the human-source counterpart to the bovine type IV SKUs (cat. 148 atelocollagen for 3D gel; cat. 231 native type IV bovine), and the type IV partner in the human-collagen autoimmunity antigen panel alongside cat. 288 (type I), cat. 210 (type II), cat. 889 (type III) and cat. 436 (type V).
SPECIFICATIONS
Catalogue number: 425
Amount: 0.1 mg
Source: Human placenta (HBsAg / HCV / HIV-1 / HIV-2 negative)
Purity: Human collagen type IV 95%. Other human collagens <5%. Non-collagen proteins <0.5%
Format: Lyophilized, salt-free
Purification: Partial pepsin digestion under acidic conditions; differential salt precipitation
STORAGE / RECONSTITUTION
Dissolve in 0.5 M acetic acid, pH 2.5.
Shipping at ambient temperature. Long-term storage: 2 years at -20 °C or lower.
APPLICATIONS
- Human type IV collagen ELISA, immunoblot and dot-blot reference standard — defined human-source antigen for quantifying type IV collagen, its NC1 domain and degradation fragments in serum, urine and tissue lysates from glomerular, diabetic-nephropathy, fibrosis, vascular and Alport-syndrome cohorts.
- Antigen for anti-human-collagen-IV antibody production — pure human-source immunogen for raising clinical-grade anti-type-IV polyclonal or monoclonal antibodies.
- Anti-GBM (glomerular basement membrane) autoantibody serological assay — the human type IV collagen antigen for screening anti-NC1 / anti-a3 autoreactivity in Goodpasture syndrome and other anti-GBM-driven glomerulonephritides.
- Coating substrate for endothelial-cell culture — primary HUVECs, human cerebral microvascular endothelial cells and induced-pluripotent-stem-cell-derived endothelial cells maintain a more native basal phenotype on basement-membrane collagen IV than on type I.
- Coating substrate for podocyte, glomerular and tubular epithelial culture — basement-membrane collagen IV is the physiological substrate for the kidney filter. cat. 425 is the species-matched human standard for primary podocyte expansion, immortalised podocyte lines and ESC / iPSC-derived kidney organoid work.
- Coating substrate for liver progenitor and hepatocyte culture — human type IV collagen supports the differentiation of liver progenitor cells, with biophysical / stiffness combinations on collagen-IV-functionalised polyacrylamide arrays driving distinct hepatocyte-vs-cholangiocyte lineage outcomes.
- ECM-microarray substrate for high-throughput differentiation, drug-response and traction-force studies — cat. 425 has been used as one of the matrix-context conditions in conjugated ECM-on-polyacrylamide microarrays at coating concentrations around 250 µg/mL, in workflows mapping cell-fate decisions, lung-adenocarcinoma drug responses and traction-force correlates of phenotype.
- Coating substrate for adipose-derived stem cell (ASC) adipogenic culture — type IV collagen supports human ASC attachment and adipogenic differentiation in 2D coatings and as a component of biomimetic encapsulating hydrogel scaffolds for tissue-engineering work.
- Monocyte-polarisation studies — type IV collagen has been used to direct THP1 monocyte differentiation toward defined macrophage states, including in autoreactive M1 macrophage protocols where cat. 425 served as the human-source type IV collagen component.
- Rescue and anoikis-protection assays — exogenous type IV collagen has been used as a survival substrate for cells driven into anoikis by basement-membrane detachment, including in ovarian-cancer Notch3 / COL4A2 upregulation models.
- Endothelial-cell rescue substrate in vascular-disease cell models — type IV collagen is one of the matrix substrates that can restore normal spheroid morphology in genetically disrupted endothelial cells (paired with fibronectin fragment cat. 175 in domain-mapping rescue experiments).
- Substrate for basement-membrane assembly and network-formation biophysics — defined-purity human type IV collagen for in-vitro reconstitution of the type IV / laminin / nidogen / perlecan basement-membrane scaffold, AFM imaging of the chloride-stabilised triple-helix-network, and structural / chloride-dependent assembly studies of NC1 hexamer and 7S tetramer junctions.
- Antigen panel for human-collagen autoimmunity research — paired with cat. 288 (type I), cat. 210 (type II), cat. 889 (type III) and cat. 436 (type V), cat. 425 completes the human-collagen serology array used in disc-, joint-, vascular- and connective-tissue autoimmunity screens.
- Substrate for type-IV-specific protease-activity assays — defined human substrate for MMP-2 (gelatinase A), MMP-9 (gelatinase B), MMP-7, cathepsin and tumour-derived basement-membrane-degrading protease cleavage assays in cancer-invasion and metastasis research.
APPLICATION HIGHLIGHTS — HIGH-VALUE RESEARCH NICHES
- Species-matched human basement-membrane substrate for epithelial, endothelial and Schwann-cell adhesion, polarity and migration assays.
- Defined single-component alternative to complex basement-membrane extracts, giving unambiguous attribution of phenotypes to collagen IV.
- Human-source standard and coating antigen for collagen-IV ELISA and for anti-GBM (Goodpasture) autoantibody screening.
- Ligand for integrin a1ß1, a2ß1, a3ß1 and aVß3 binding studies and for integrin-blocking experiments.
- Substrate for basement-membrane remodelling and matrix-metalloproteinase degradation assays in tumour-invasion research.
- Immunogen and QC antigen for anti-human collagen type IV antibody development.
REFERENCES
(1) Kuzan A., Chwilkowska A., Maksymowicz K., Bronowicka-Szydelko A., Stach K., Pezowicz C., Gamian A. (2018) — Advanced glycation end products as a source of artifacts in immunoenzymatic methods. Glycoconjugate Journal 35(1): 95–103. doi:10.1007/s10719-017-9805-4.
The authors used cat. 425 as the human type IV collagen reference standard in an ELISA / immunoblot panel covering 103 human aorta fragments — type I, III and IV collagens, elastin and AGE content. Establishes how AGE accumulation on collagens biases immunoenzymatic readouts, and how a defined human-collagen-IV reference is essential to correct for these artefacts when analysing vascular basement-membrane glycation in diabetic and atherosclerotic disease.
(2) Pal S., Konkimalla V.B. (2016) — Data on sulforaphane treatment mediated suppression of autoreactive, inflammatory M1 macrophages. Data in Brief. doi:10.1016/j.dib.2016.03.105.
The authors used cat. 425 to polarise THP1 monocytes toward an autoreactive M1 macrophage phenotype, supporting collagen-IV / autoreactive-macrophage interaction as a druggable target in autoimmune-inflammation drug discovery. cat. 425 supplied the species-matched human type IV antigen needed for human-cell-based autoreactivity readouts.
