Description
Native-conformation human collagen type I purified from human placenta — the rare tissue source that allows a true species-matched type I collagen for human cell-biology workflows. Purification proceeds by partial pepsin digestion under acidic conditions, differential salt precipitation and DEAE ion-exchange chromatography, yielding a lyophilized product of =90 % type I purity with <10 % type III and <1 % other collagens or non-collagen contaminants. Each lot is QC-controlled by SDS-PAGE and by ELISA with type-specific anti-collagen antibodies; donor is screened negative for HBsAg, HCV and HIV-1/HIV-2.
Because the product is human-source and partially pepsin-treated (telopeptides only partially trimmed), cat. 288 occupies a specific position in the YO Proteins catalogue: it preserves the immunological reactivity needed for human-collagen ELISA, immunoblot and anti-collagen antibody work, but is not intended for 3D gel formation. For sterile bovine, mouse, rat or porcine atelocollagens optimised for 3D gel use, see cat. 437, 176, 602, 130 / 882. For full-length and fragment human plasma fibronectin, see cat. 663 / 175 / 815.
The product has been used in eleven published studies as a human-source coating substrate (10 µg/cm² to 40 µg/mL), an ELISA / dot-blot reference standard for human type I collagen, an antigen in autoimmunity screens, and a rescue / loss-of-function reagent in cancer-invasion and stem-cell-derivation work. The Cited use cases section below summarises each in turn.
Specifications
Catalog number: 288
Amount: 0.1 mg
Source: Human placenta (HBsAg / HCV / HIV-1 / HIV-2 negative)
Purity: Human collagen type I =90 %, human collagen type III <10 %, other human collagens & non-collagen proteins <1 %. QC by SDS-PAGE and ELISA with type-specific antibodies
Format: Lyophilized
Purification: Partial pepsin digestion under acidic conditions, differential salt precipitation, DEAE ion-exchange chromatography
Reconstitution
Dissolve in 0.1 M acetic acid, pH 3.0. Recommended working concentration depends on the application — see typical coating / standard concentrations under Applications.
Storage
Shipping at ambient temperature.
Long-term storage: 2 years at -20 °C or lower.
Applications
- **Human type I collagen ELISA & dot-blot standard** — defined human-source reference for quantification of type I collagen in cell lysates, conditioned media, tissue digests and biofluids. Especially valuable where a species-matched human standard is required to avoid epitope mis-detection seen with bovine or rodent substitutes.
- **Antigen for anti-human collagen type I antibody production** — pure human-source immunogen for rabbit, mouse or chicken immunisation protocols.
- **Culture-surface coating for human cell adhesion, migration and rescue assays** — typical coating range 10 ng/mL (sensitive primary models) to 40 µg/mL or 10 µg/cm² (robust attachment of fibroblasts, MSCs, cancer lines).
- **Substrate for collagen-glycation studies** — native human collagen I can be glycated in vitro (mannose, rhamnose, arabinose, glucose) to investigate AGE-driven stem-cell differentiation and ECM-receptor interactions.
- **Cell-stiffness / topography substrates** — incorporated into hydrogels of defined Young's modulus, or aligned with magnetic colloidal scaffolds, to decouple stiffness from fibrillar topography in cell-mechanobiology experiments.
- **Antigen panel for autoimmunity / autoantibody screens** — human-source type I collagen for serum-IgG / IgM autoantibody ELISA in intervertebral-disc, vascular and inflammatory pathology.
- **Rescue / loss-of-function reagent** — supplemented to medium or surface-coated to test whether a phenotype downstream of an ECM regulator (e.g. PERK, CREB3L1) can be restored by exogenous type I collagen.
APPLICATION PROTOCOL EXAMPLE — Cell attachment assay (HeLa)
A representative protocol used in cell-attachment screens with cat. 288:
1. Detach HeLa cells with non-enzymatic dissociation buffer.
2. Reseed onto 96-well polystyrene plates pre-coated with 40 µg/mL human collagen type I (cat. 288, in 0.02 M acetic acid; coat 1 h at room temperature, rinse with PBS).
3. Incubate cells for 30 min at 37 °C.
4. Wash non-adherent cells away with DMEM.
5. Quantify attached cells by the crystal-violet spectrophotometric method (A595 after acetic-acid elution).
(Adapted from Rui et al. 2017, Cell Physiol Biochem — see Cited use case 9.)
CITED USE CASES
(1) Brokopp C.E. et al. (2011) — Fibroblast activation protein is induced by inflammation and degrades type I collagen in thin-cap fibroatheromata. European Heart Journal 32(21): 2713–2722. doi:10.1093/eurheartj/ehq519.
Used cat. 288 as the full-length native human type I collagen substrate (100 ng/mL) to demonstrate that recombinant human Fibroblast Activation Protein (FAP, 200 nM) specifically cleaves type I collagen — establishing FAP as a driver of fibrous-cap thinning in vulnerable atherosclerotic plaque. cat. 288 served as the gold-standard human substrate for FAP/cathepsin protease-activity assays.
(2) Capossela S. et al. (2014) — Degenerated human intervertebral discs contain autoantibodies against extracellular matrix proteins. European Cells and Materials 27: 251–263. doi:10.22203/eCM.v027a18.
Used cat. 288 as the human collagen type I ELISA / immunoblot antigen (alongside cat. 889 type III and cat. 436 type V) to screen serum and disc-tissue extracts of degenerated-disc patients for anti-ECM autoantibodies. Demonstrated for the first time the existence of disc-targeted autoimmunity. cat. 288 is the validated human type I antigen for the assay.
(3) 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. Cosmetics 2(2): 146–161. doi:10.3390/cosmetics2020146.
Used cat. 288 as the human type I collagen reference standard for dot-blot / ELISA quantification of fibroblast-secreted type I, III and V collagens and HSP-27 / 47 / 70 / 90 — assessing the anti-photoaging effect of nicotinamide derivatives on UVA-radiated dermal fibroblasts.
(4) Kuzan A. et al. (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.
Used cat. 288 as the human collagen type I 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 I reference is essential to correct for these artefacts.
(5) Philips N. et al. (2012) — Beneficial regulation of fibrillar collagens, heat shock protein-47, elastin fibre components, transforming growth factor-ß1, VEGF and oxidative stress effects by copper in dermal fibroblasts. Connective Tissue Research, 1–7.
Used cat. 288 as the human collagen type I ELISA / dot-blot standard in a study of copper-driven regulation of fibrillar collagen, HSP-47, elastin, TGF-ß1 and VEGF in dermal fibroblasts.
(6) Mogami H., Keller P.W., Shi H., Word R.A. (2014) — Effect of thrombin on human amnion mesenchymal cells, mouse fetal membranes, and preterm birth. Journal of Biological Chemistry 289(19): 13295–13307.
Used cat. 288 as the human type I collagen reference for ELISA quantification of collagen synthesis by amnion mesenchymal cells, in a study showing that thrombin increases MMPs and COX-2 via PAR-1 / TLR4 and drives preterm birth in mice.
(7) Duruksu G., Aciksari A. (2018) — Guiding the differentiation direction of pancreatic islet-derived stem cells by glycated collagen. Stem Cells International 2018: 6143081. doi:10.1155/2018/6143081.
Used cat. 288 as a culture-plate coating at 10 µg/cm², either as native human type I collagen or after in-vitro glycation with mannose, rhamnose, arabinose and glucose. Rat pancreatic-islet-derived mesenchymal stem cells (PI-MSCs) were seeded onto the coated surfaces; native vs. glycated collagen I produced markedly different differentiation outcomes, demonstrating that the AGE state of the matrix is a directional cue for PI-MSC fate. Establishes cat. 288 as the standard human substrate for glycation-and-stem-cell-differentiation studies.
(8) Paul C.D., Hruska A., Staunton J.R., Burr H.A., Kim J., Jiang N., Tanner K. (2017) — Decoupling cellular response to topography and stiffness in three dimensions. bioRxiv 232066. doi:10.1101/232066.
Used cat. 288 as the human type I collagen incorporated into 3D amorphous hydrogels with fibrillar architecture built by magnetic-colloidal self-assembly. Cell topography of human foreskin fibroblasts (HFF) and U87 human glioblastoma cells was assessed independently of bulk stiffness — establishing cat. 288 as a defined human-collagen building block for 3D mechanobiology constructs.
(9) Rui Y.-N. et al. (2017) — The intracranial aneurysm gene THSD1 connects endosome dynamics to nascent focal adhesion assembly. Cellular Physiology and Biochemistry 43(6): 2200–2211. doi:10.1159/000484298.
Used cat. 288 in HeLa cell-attachment assays (40 µg/mL coating, 30 min adhesion, crystal-violet quantification) to demonstrate that THSD1 — the intracranial-aneurysm susceptibility gene — links endosome dynamics to nascent focal-adhesion assembly on type I collagen. cat. 288 is the human-source substrate that anchors the assay to disease-relevant ECM biology.
(10) Dumevska B., Chami O., McKernan R., Goel D., Schmidt U. (2016) — Derivation of Genea052 human embryonic stem cell line. Stem Cell Research 16(2): 327–330. doi:10.1016/j.scr.2015.12.043.
Used cat. 288 at 10 ng/mL as the gentle coating substrate for plating the zona pellucida of a blastocyst-stage human embryo, supporting inner-cell-mass outgrowth on inactivated human feeders and the subsequent derivation of the Genea052 hES cell line (46,XY). Demonstrates cat. 288's suitability at very low coating densities for delicate primary human-embryo work.
(11) Feng Y.-X. et al. (2017) — Cancer-specific PERK signaling drives invasion and metastasis through CREB3L1. Nature Communications 8: 1079. doi:10.1038/s41467-017-01052-y.
Used cat. 288 in collagen type I rescue experiments to test whether exogenous human collagen I restores phenotypes downstream of PERK / CREB3L1 in mesenchymal triple-negative breast-cancer cells. The study identifies CREB3L1 as the essential transcription factor through which PERK drives invasion and metastasis — and establishes cat. 288 as the rescue-reagent of choice for cancer-cell collagen-loss assays.
