| Size | Price | Stock | Qty |
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| 10g |
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| 50g |
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| Targets |
Integrins (cell surface receptors that mediate cell-extracellular matrix adhesion). Type III collagen interacts with integrins to regulate cellular functions such as adhesion, migration, proliferation, and differentiation. The specific integrin subtypes that bind to rhCOLIII include alpha1beta1, alpha2beta1, alpha10beta1, and alpha11beta1.
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| ln Vitro |
In vitro, recombinant humanized type III collagen (rhCOLIII) inhibits the proliferation, migration, and invasion of breast cancer cells. The protein promotes skin extracellular matrix (ECM) regeneration and improves the cell microenvironment by providing a natural scaffold that supports cell growth and migration. The specific IC₅0 for inhibition of cancer cell proliferation is not provided in the available data.
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| ln Vivo |
A general in vivo activity of Recombinant Humanized Type III Collagen is the promotion of wound healing and skin regeneration. In animal models of dermal wounds, application of rhCOLIII accelerates wound closure, increases granulation tissue formation, and enhances re-epithelialization. The protein also has anti-tumor effects in breast cancer xenograft models, inhibiting tumor growth, migration, and invasion.
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| Enzyme Assay |
A general cell-free protocol for assessing integrin binding: A solid-phase binding assay is used. Recombinant integrin protein (e.g., alpha1beta1 or alpha2beta1, 10 ug/mL in 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM MnCl2) is coated onto 96-well microtiter plates overnight at 4degC. The wells are blocked with 3% BSA in PBS for 2 hours. Varying concentrations (0.1, 0.5, 1, 5, 10, 20, 50, 100 ug/mL) of Recombinant Humanized Type III Collagen (rhCOLIII) are added to the wells and incubated for 2 hours at room temperature. Bound collagen is detected using an anti-collagen III antibody conjugated to HRP, followed by the addition of a TMB substrate. The absorbance is measured at 450 nm. The Kd for the integrin-collagen interaction is calculated from the saturation binding curve. For competition assays, a fixed concentration of rhCOLIII (10 ug/mL) is incubated with varying concentrations of soluble integrin (0.1-100 ug/mL) or with an anti-integrin blocking antibody.
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| Cell Assay |
A general cellular protocol for assessing the effects of rhCOLIII on breast cancer cells: Human breast cancer cell lines (e.g., MDA-MB-231, MCF-7, or 4T1 cells) are seeded in 96-well plates at 5,000 cells/well in DMEM containing 10% FBS. After 24 hours, the cells are treated with various concentrations of rhCOLIII (10, 25, 50, 100, 200 ug/mL) for 24, 48, and 72 hours. Cell viability is assessed using the MTT assay. For proliferation assays, cells are incubated with BrdU for 2 hours, and BrdU incorporation is measured by ELISA. For migration and invasion assays, Transwell chambers with 8-um pores are used. For migration, cells (1×10⁵) in serum-free DMEM are placed in the upper chamber. The lower chamber contains DMEM with 10% FBS as a chemoattractant. rhCOLIII (50, 100, 200 ug/mL) is added to both chambers. For invasion assays, the upper chamber is coated with Matrigel (50 uL of 1 mg/mL). After 24 hours, the cells on the upper surface of the membrane are removed, and the cells on the lower surface are fixed, stained with crystal violet, and counted.
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| Animal Protocol |
A general animal protocol for evaluating wound healing efficacy: Female Sprague-Dawley rats (200-250 g) are used. The animals are anesthetized, and the dorsal fur is shaved. A full-thickness excisional wound (1.5 cm × 1.5 cm) is created on the dorsum using sterile scissors and forceps. Recombinant Humanized Type III Collagen is formulated in sterile PBS as a solution (1, 5, and 10 mg/mL) or as a hydrogel. The test article (100 uL of solution or a hydrogel patch) is applied topically to the wound site once daily for 14 days. A vehicle control group (PBS) and a positive control group (e.g., a commercial wound dressing) are included. Wound area is measured by tracing the wound margin on days 0, 3, 5, 7, 10, and 14, and the percentage of wound closure is calculated. On day 14, the rats are euthanized, and the wound tissue is excised. One part of the tissue is fixed in 10% formalin, embedded in paraffin, sectioned, and stained with H&E and Masson‘s trichrome for histopathological examination to assess re-epithelialization, granulation tissue formation, and collagen deposition. Another part is used for hydroxyproline assay (a marker of collagen content).
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| ADME/Pharmacokinetics |
A general pharmacokinetic protocol is not applicable for a protein. Recombinant Humanized Type III Collagen is a large protein (80-100 kDa) that is not intended for systemic absorption. In topical applications, the protein is expected to remain at the application site and be degraded locally. If administered intravenously, it would be rapidly cleared by the liver and kidneys. A general protocol for assessing degradation: The protein is incubated in simulated wound fluid or in a solution containing proteases (e.g., collagenase, MMP-1, MMP-8, MMP-13) at 37degC. The degradation products are analyzed by SDS-PAGE and Western blot at various time points.
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| Toxicity/Toxicokinetics |
General toxicity protocol for topical application: A dermal irritation/corrosion test is performed in rabbits according to OECD Guideline 404. Approximately 0.5 g of Recombinant Humanized Type III Collagen (as a gel or solution) is applied to a small area (approximately 1 cm2) of intact and abraded skin on the dorsum of three rabbits. The application site is covered with a gauze patch and secured with non-irritating tape. After 4 hours of exposure, the patches are removed, and the skin is cleaned. The skin is examined for erythema and edema at 1, 24, 48, and 72 hours after patch removal. A general systemic toxicity protocol for intravenous administration: A single-dose toxicity study is performed in mice. Recombinant Humanized Type III Collagen is administered via intravenous (IV) injection at doses of 50, 150, and 500 mg/kg. The animals are observed for 14 days for clinical signs. At the end of the observation period, the animals are euthanized, and a necropsy is performed. The major organs are collected for histopathological examination.
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| References |
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| Additional Infomation |
Recombinant Humanized Type III Collagen (80-100kDa) is a recombinant humanized type III collagen produced in a microbial expression system. It has an apparent molecular weight of 80-100 kDa as determined by SDS-PAGE. The protein is soluble in water and forms a clear solution. The pI is 6.59. The product is supplied as a sterile lyophilized powder. Recombinant Humanized Type III Collagen has biological functions including promoting skin extracellular matrix regeneration, improving the cell microenvironment, inhibiting the proliferation, migration, and invasion of breast cancer cells, and functioning in cell adhesion, migration, proliferation, and differentiation through interaction with integrins. It is a research-grade protein for cell culture studies, tissue engineering, and cancer research.
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| Appearance |
Solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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