Product DescriptionType I bovine collagen, lyophilized fibrous sheet, Catalog Number 5164, is extracted from bovine flexor tendon with the raw material sourced from closed/controlled herds of animals. Because the collagen is extracted from tendon, the material is more naturally crosslinked. The collagen is polymericinsolublecollagen. The manufacturing processes comply with stringent quality standards that have proven to yield a high quality product with lot-to-lot consistency. This product has a purity of >96% with Type II and Type III collagens not detectable.This product is supplied as a lyophilized fibrous sheet in a 5 gram package size. Bioburden and endotoxin levels are tested – this product is not considered sterile.This collagen product is naturally cross-linked yielding a robust material for applications which require structure and strength. This product can be readily prepared into such forms as tissue scaffolds, foams, sponges, suspensions, coatings, putties, films and sheets butdoes not form hydrogels. Using typical cross-linking methods, this material can be tuned for optimal in vivo resorption. This collagen product is ideal for tissue engineering applications and uses with inorganic and biomaterials.The product is provided in user-friendly packaging for use and storage. Avoid extended open-air exposure to environment since this material is hygroscopic.Parameter, Testing, and MethodInsoluble Type I Collagen Sheet #5164FormLyophilized FibrousSheetPackage Size5 gramsStorage TemperatureRoomtemperatureShelf LifeMinimum of 6 months from date of receiptCollagen Purity>96%Endotoxin - LAL<20.0 EU/mLBioburden<200 cfu/gramCell Attachment AssayPassSourceBovineFlexor TendonAmino Acid AnalysisCharacteristicDirections for UseDownload the full PDF versionor continue reading below:Note: The following procedure is based on preparationof 1 gram of collagen in 100 ml to initially prepare a 10 mg/ml collagen suspension. Smaller quantities and volumes may be used but the same ratios of collagen and solutions should be used.Weigh out 1 gram of collagen fibrous sheet.Cut the collagen sheet into approximately 0.5 cm2 pieces.Reconstitute 1 gram of collagen pieces with 50 ml of cold purified water (50% of the final volume).Stir the collagen with the water continuously mixing for a minimum of 15 minutes until the collagen is fully wetted and the suspension appears to be a semi-solution with some lumps.Add 50 ml of cold 0.02 M HCl to the collagen mixture and stir for a minimum of 10 minutes. This will yield a collagen concentration of 10 mg/ml with the suspension continuing to appear as a semi-solution with lumps.Measure the pH – the mixture should have a pH of 2 to 3.Using Waring stick blender or equivalent, homogenize the collagen mixture for a minimum of 15 minute at a high speed ensuring that the collagen is fully homogenized. Ensure that the temperature of mixture does exceed 24°C. Upon completion, there should be very few visual solids in the viscous suspension. Air bubbles with be prevalent.To remove air bubbles, stir the solution on a stir plate and pull a vacuum on the suspension. This will remove the air bubbles.At this point if a collagen concentration of less than 10 mg/ml is desired, the collagen can be diluted with 0.01 M HCl.Product Q & AIs there DNA in the collagen products?We completed a study to show that DNA is completely destroyed at pH 2, and demonstrated that our collagen products do not contain DNA.How is the Amino Acid Analysis performed?The collagen is fully hydrolyzed. The amino acid analysis is done using the Waters AccQ-Tag derivatization method.During the acid hydrolysis step, asparagine (N) is converted to aspartic acid (D) and glutamine (Q) is converted to glutamic acid (E). Tryptophan (W), if present, is destroyed during acid hydrolysis. Experimentally, one can determine the picomoles (pmol) of each amino acid per injected detected using amino acid standards.For the concentration determination, the total number of pmol of each amino acid is summed to get the total pmol of the 18 amino acids detected. The total pmol amino acids is divided by the theoretical number of amino acid residues in collagen based on the published sequence. The result is the pmol of collagen injected. The result is then multiplied by the dilution and 300,000 is used as the collagen molecular weight to get to mg/mL. The molecular weight of collagen is not well agreed upon.Can you dilute collagen with 1X PBS for 2D coatings?Diluting with 1X PBS (rather than water or 0.01 N HCl) would have an effect for coating purposes. It would change the pH of the diluted collagen solution from acid to neutral pH. The pH change will transform the collagen molecules from a molecular form to a fibrillar form; and then the nature of coating surface will be changed from a monomeric coating to a fibrillar coating.What antibodies do you recommend for your Type I collagen products?We use thefollowing antibodies from SouthernBiotech:1. 1310-02 – Goat Anti-Type I Collagen-FITC2. 1310-08 – Goat Anti-Type I Collagen-BIOT3. 7100-05 – Streptavidin-HRPWhat is the thickness of the ‘molecular collagen’ attaching to the polystyrene surface of cultureware?The major collagen molecular species in our Type I collagen products are monomers (approx. 70%), but there are dimers, trimers and a few percentages of oligomers too (approx. 30%) with some minor amounts of collagen fragments. The collagen monomer is a rod shaped molecule with 300 nm in length and 1.5 nm in diameter. The dimer, trimer and oligomer are 600 nm, 900nm and even longer in length respectively. According to the coating procedures, the collagen molecules are attached to the charged polystyrene surface randomly by charge or affinity in acid conditions during the 1-2 hrs incubation period at 37°C, and any unattached materials are removed by aspiration and rinsing. Therefore, the coated surface is a single layer of collagen monomer, dimer, trimer and oligomer mixtures.The thickness of the mono-molecular layer is dependent on how those molecules are attached on the surface. The coating density thickness would generally be characterized as a 1 molecule thickness which could be ranging from a few nanometers to a few hundred nanometers with the whole surface being covered by collagen.What is the charge of your collagen products?The net charge of Type I collagen products’ (PureCol®, Bovine Collagen and VitroCol®, Human Collagen) molecule is directly related to the pH. At an acidic pH, the amino acids (zwitterions) along the collagen molecule are positively charged, making the entire collagen molecule positive. At the isoelectric point (or zone) of collagen, around pH 7-8, the amino acids along the collagen molecule are positively and negatively charged, making the net charge of the collagen molecule close to zero. At a basic pH, the amino acids along the collagen molecule were negatively charged, making the entire collagen molecule negative.Further, the nature of the charge of the collagen coating surface will be dependent on the type of coating applied. For a monomeric collagen coatings when the collagen is applied under an acidic pH condition, the surface is positively charged. If the surface is rinsed with pH neutral buffer or media then it will change the charge of the collagen surface net charge close to zero. For a 3D gel coating, the collagen prepared under neutral pH; the net charge of the collagen surface is close to zero.What is the monomer to oligomer ratio in your collagen products?Using rotary shadowing technique under transmission electron microscopy, it was found that our collagen, on average, consists of approximately 80% monomers, 13% dimers, trimers, and oligomers with the remaining 7% collagen fragments.Is pepsin treated collagen still MMP-Sensitive?Yes.The collagen molecule in PureCol, Nutragen, VitroCol, and all of our other Atelo collagen products were prepared from native collagen matrix by pepsin treatment under controlled conditions to remove the non-helical portion, telo-peptides, only and the helical portion is intact. In this case, the enzymatic active sites for MMP (Matrix Metalloproteinase), such as for Mammalian Collagenase Matrix Metalloproteinase 8 (MMP-8), on the molecule was preserved.These pepsin treated collagen products should behave as native intact collagen.Is there TGF Beta in the collagen products?TGF beta would have been digested with the pepsin enzymatic digestion step. It was undetectable by SDS PAGE silver stain as well. We didn’t do any specific measurements by ELISA however but presences of TGF betais not anticipated.How do you determine protein concentration?We primarily use the Biuret method, but we also use BCA, AAA, and hydroxyl-proline assays.Is my collagen OK if it was frozen, or left out at room temperature?- Collagen solutions that are frozen tend to have issues forming 3D hydrogels, and will likely not work. The solutions should still be good for 2D coatings.- Collagen solutions that are left out at room temperature for extended periods of time may show signs of degradation, which will affect the formation of 3D hydrogels. It is likely still fine for 2D coatings.Our recommendation is this: If you are using the product directly for a publication, we highly suggest buying a new bottle if the one you have was compromised. Product ReferencesReferences for Insoluble Collagen:Ahmed, Raju, Monjurul Haq, and Byung-Soo Chun. "Characterization of marine derived collagen extracted from the by-products of bigeye tuna (Thunnus obesus)."International journal of biological macromolecules(2019).Divakar, Prajan, Kaiyang Yin, and Ulrike GK Wegst. "Values and property charts for anisotropic freeze-cast collagen scaffolds for tissue regeneration."Data in brief22 (2019): 502-507.Divakar, Prajan, Kaiyang Yin, and Ulrike GK Wegst. "Anisotropic freeze-cast collagen scaffolds for tissue regeneration: How processing conditions affect structure and properties in the dry and fully hydrated states."Journal of the mechanical behavior of biomedical materials90 (2019): 350-364.Product Certificate of AnalysisNo result for .Product Videos30+ Type I Collagen OptionsVideoSee More >Why 3D MattersVideoSee More > See More Safety and DocumentationSafety Data SheetCertificate of OriginProduct DisclaimerThis product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

美国Advanced BioMatrix是3D组织培养、细胞检测和细胞增殖等领域实验解决方案的佼佼者。Advanced BioMatrix在分离、纯化、冻干、细胞培养和蛋白检测、多肽粘附、附着因子、基质硬度和其他3D matrix 产品开发方面有着丰富的经验。Advanced BioMatrix的研发经验和专业知识确保其产品可达到最佳质量，并保证产品之间一致性，方便研究客户使用。

以下为Advanced BioMatrix 3D Matrices 产品竞争优势：
1. 提供高纯度和成分确定的胞外基质；
2. 超过1000余篇文献引用PureCol产品，品质非常均一；
3. 在3D培养基领域可提供最全面的产品线；
4. 唯一可提供特异性刚性有机硅基板的公司（CytoSoft）;
5. 唯一可提供可溶性丝纤蛋白的供应商（可运用于多种3D培养）；
6. 如果客户首次接触3D胶原凝胶，Advanced BioMatrix还是唯一的预制胶原蛋白(PureCol EZ Gel)供应商。

以下产品为Advanced BioMatrix全球畅销品：
1. PureCol 牛源I型胶原蛋白 3 mg/ml #5005-100ML
2. Nutragen牛源I型胶原蛋白 6 mg/ml #5005-100ML
3. FibriCol 牛源I型胶原蛋白 10 mg/ml #5133-20ML
4. VitroCol 人源I型胶原蛋白 #5007-20ML
5. 弹性蛋白原 #5052-1MG
6. ECM Select Array kit Ultra-36 #5170-1EA
7. CytoSoft（刚性可变的基底，Advanced BioMatrix最新添加产品5190-7EA）
8. 人III型胶原蛋白 #5021-10MG
9. 人IV型胶原蛋白 #5022-5MG
10. Silk Fibroin溶液 #5154-20ML
11. Fibronectin #5080-5MG
12. Vitronectin #5051-0.1MG

Advanced BioMatrix最畅销的产品是PureCol（5005-100ML）。目前已有上千发表的论文使用这款产品。（Nutragen 5010、fibricol 5133和PureCol的物质成分一样，仅仅是浓度不同）浓度越高，胶原蛋白硬度越强。