Product Description
Overview
The HyStem-IRG QuickSet Kit is composed of Glycosil® (thiolmodified hyaluronic acid), Gelin-S® (thiol-modified gelatin), PEGCure (PEG-norbornene) and Irgacure 2959 Photoinitiator for photoinitiation. A transparent hydrogel forms after contents are mixed and exposed to UV light (365 nm). All vials are packaged as sterile lyophilized solids that are blanketed by argon and under a slight vacuum.
As one of the first hyaluronic acid-based light-controlled hydrogels, the HyStem®-IRGQuickSet Kit supports 3D cell culture for use in tissue engineering and small scale bioprinting applications. The HyStem®-IRGQuickSet Kitprovides increased temporal and spatial control making it ideal for micro-scale bioprinting. Gelation occurs after 30-60 seconds when exposed to ultraviolet light as compared to ~30 minutes using other hydrogel formulations.
Features
- Enables faster, flexible gelation times
- Controlled by UV light
- Optimized for micro-scale bioprinting applications
- Maintains standard characteristics of other HyStem hydrogels
The HyStem®UV QuickSet kit is composed of HyStem (thiol-modified hyaluronic acid), Gelin-S®(thiol-modified gelatin), UVlink™ (PEG-norbornene), Irgacure 2959 for photoinitiation, and DG water. A transparent hydrogel forms when contents are mixed and exposed to UV light. All vials are packaged as sterile lyophilized solids that are blanketed by argon and under a slight vacuum.
Directions for Use
INSTRUCTIONS FOR USE (Download the full directions for use PDF here)
The Irgacure solution is prepared by dissolving the lyophilized solid in DG Water. The Glycosil, Gelin-S, and PEGCure are reconstituted with the Photoinitiator solution. A 7.5 mL hydrogel at 1% (w/v) solution is produced when all reconstituted materials are mixed.
HyStem hydrogels (3 x 2.5 mL = 7.5 mL) should be prepared as follows:
- Allow Glycosil, Gelin-S, PEGCure, irgacure, and DG Water vials to come to room temprerature.
- Under aseptic conditions, use a syringe to add 10.0 mL of DG Water to the Irgacure Photoinitiator vial. Shake or vortex the vial at 37°C for 30 minutes or until fully dissolved.
- Add 1.0 mL of the reconstituted Photoinitiator to the Glycosil vial. Add 1.0 mL of the Photoinitiator solution to the Gelin-S vial.
- Place both vials horizontally on a rocker or shaker. Shake vials at 37°C for 30 minutes or until fully dissolved. It may take up to 60 minutes for the solids to fully dissolve.
Solutions should be clear and slightly viscous.
- Add 0.5 mL of Photoinitiator to the PEGCure vial. Place on a shaker and mix at 37°C for approximately 10 minutes.
- Combine the Glycosil, Gelin-S, and PEGCure solutions and mix well.
- Pipette solution into desired format (i.e. 96 well plate). Using a hand-held UV light source, expose the gel to the UV light (wavelength 365nm) until the gel reaches the desired stiffness. Gelation will occur between 15 seconds and 1 minute. Results may vary depending on the UV source manufacturer and/or plate design
Note: Gelation time and gel stiffness can be adjusted by varying the concentration of Glycosil, GelinS, or UVlink.
Note: Each kit component has been manufactured under aseptic conditions and tested for bacteria and fungus.
Product Q & A
Globular particles less than 75 kDa should be able to freely diffuse through a HyStem hydrogel.
When reconstituted using DG water, the pH of each HyStem component will be approximately 7.4-7.6.
One year from the date of receipt, if stored properly.
Any sterile, deionized, degassed water can be substituted for reconstitution. However, in order to ensure accurate and predictable dissolution and gelation times, our DG Water is highly recommended, as it is degassed, blanketed in argon, and has undergone validation testing with each HyStem component.
Gelin-S provides cellular attachment sites when incorporated in the hydrogel. Gelin-S is thiol-modified, denatured collagen I, derived from either bovine or porcine sources. Gelin-S is included in all HyStem-C and HyStem-HP kits.
Gelin-S has been thiol-modified in the same manner as the hyaluronan in Glycosil (or Heprasil), so that it covalently crosslinks with the Extralink in the HyStem hydrogels.
Yes. Peptides that contain a cysteine residue can be used. The cysteine residue must be present for the peptide to be covalently bonded to the hydrogel substrate.
Yes. ECM proteins, such as laminin, collagen, fibronectin, or vitronectin can be non-covalently incorporated into the hydrogel prior to crosslinking.
HyStem hydrogels and sponges differ in hydration and homogeneity. HyStem sponges are typically polymerized hydrogels that are subsequently freeze-dried. The resulting sponge is a fibrous, mesh network with pores and niches that enable cells to infiltrate and adhere. A true HyStem hydrogel is an encapsulating liquid that polymerizes around suspended cells in culture.
No. The compliance of the hydrogels is set by the amount of Extralink crosslinker added, the concentration of Glycosil (or Heprasil) and Gelin-S used, and the ratio of Glycosil (or Heprasil) to Gelin-S. Once this chemical structure of the hydrogel is fixed, it is not altered by prolonged exposure to cell culture medium.
HyStem sponges can be terminally sterilized by E-beam. HyStem hydrogels have not yet been validated for use with E-beam sterilization methods. HyStem hydrogels are not terminally sterilized by gamma irradiation.
Gelation time is affected by multiple aspects of the gel’s composition.One way to change the gelation time of a hydrogel is to vary the amount of crosslinker used. Gels with a lower amount of Extralink crosslinker will have a longer gelation time than those with a higher amount of crosslinker. Changing the amount of crosslinker will produce slight changes in gelation time.Gelation time can be dramatically changed by varying the Glycosil (or Heprasil) and Gelin-S concentrations. Concentrated solutions of Glycosil (or Heprasil) and Gelin-S will create a solution with a much shorter gelation time. This can easily be done by reconstituting the components in a smaller volume of DG Water. Alternatively, diluting these components in larger volumes of DG Water will dramatically increase the total time to form the hydrogel.
HyStem Hydrogels are virtually transparent and should not interfere with microscopy.
HyStem hydrogels may generate mild inflammation as part of the body’s natural healing process in response to injury. HyStem hydrogels do not trigger immune response when used in vivo. (These products are not for human use)
HyStem is degraded in vivo by matrix metalloproteinases (collagenases) and hyaluronidases.
Trypsin, Dipase, collagenase, and hyaluronidase have been used to help detach cells from the surface or from within HyStem hydrogels.
In general, the pore size for HyStem-C and HyStem-HP hydrogels is ~17 nm.
Product Applications
General Notes for Bioprinting with HyStem®-IRG QuickSet Kit:
The following guidelines were developed for a modified Bioforce Nano eNabler™ and exact requirements and parameters will vary between platforms and applications. While other platforms have not been evaluated, any system capable of liquid handling and amenable to UV exposure should be easily adaptable for the HyStem®-IRG QuickSet matrix.
• High relative humidity (>90%) is required in the printing chamber to prevent desiccation of printed constructs.
• If printing live cells, verify that the surface patterning tool or print head is large enough to accommodate cells (approximately 50-100 µm).
• Where possible, UV/ozone treatment of printing tools to enhance hydrophilic nature of the printing channel materials can improve consistency in printing.
• The matrix is a clear liquid with minimally increased viscosity compared to water at room temperature. Upon exposure to 365nm UV light, rapid polymerization occurs resulting in a viscoelastic solid. A 4W handheld UV lamp held approximately 2 cm from the surface of the gel is sufficient to polymerize a gel in as little as 15 seconds.
• Conducting a small pilot study to determine the appropriate exposure time for a given application is strongly recommended as matrix stiffness is directly correlated to exposure time and can impact survivability, proliferation, and differentiation.
Note: The HyStem®-IRG QuickSet Kit has not been thoroughly evaluated for thick, multilayer constructs.
Product Certificate of Analysis
Safety and Documentation
Certificate of Origin
Safety Data Sheet
Product Disclaimer
This 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AdvancedBioMatrix（简称ABM） www.advancedbiomatrix.comAdvancedBioMatrix（简称ABM）是美国一家著名的生物公司，获得了AllerganInc的授权(Allergan用25年时间不断完善胶原蛋白相关的产品的生产工艺)，将Allergan的专业和技术用于蛋白生产与检测，致力于为组织工程、细胞分析及细胞增殖等研究领域提供优质稳定的产品。AdvancedBioMatrix不断丰富已有产品线，目前可为三维细胞培养提供各种胶原蛋白、纤连蛋白、玻连蛋白、水性凝胶、不同粘度与分子量的透明质酸以及低代成纤维细胞等。在美国全部产品授权Sigma销售。AdvancedBioMatrix是组织培养，细胞分析和细胞增殖三维（3D）应用的生命科学领域的领导者。我们的产品被公认为纯度，功能性和一致性的标准。我们在生产，分离，纯化，冷冻干燥，细胞培养和蛋白质测试，粘附肽，附着因子，底物刚性和其他3D矩阵产品方面拥有丰富的专业知识。我们的专业技术和知识正在被用来确保我们的产品质量最高，批次之间一致且易于为我们的研究客户使用。
美国AdvancedBioMatrix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。AdvancedBioMatrix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3Dmatrix 产品开发方面有着丰富的经验。AdvancedBioMatrix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AdvancedBioMatrix3DMatrices 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，AdvancedBioMatrix还是唯一的预制胶原蛋白(PureColEZGel)供应商；
以下产品为AdvancedBioMatrix全球畅销品：1.PureCol 牛源I型胶原蛋白 3mg/ml#5005-100ML2.Nutragen牛源I型胶原蛋白 6mg/ml#5010-50ML3.FibriCol 牛源I型胶原蛋白 10mg/ml#5133-20ML4.VitroCol 人源I型胶原蛋白 #5007-20ML5. 弹性蛋白原 #5052-1MG6.ECMSelectArraykitUltra-36#5170-1EA7.CytoSoft（刚性可变的基底，AdvancedBioMatrix最新添加产品5190-7EA）8. 人III型胶原蛋白 #5021-10MG9. 人IV型胶原蛋白 #5022-5MG10.SilkFibroin溶液 #5154-20ML11.Fibronectin#5080-5MG12.Vitronectin#5051-0.1MG
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elisa试剂盒回收率是反应待测物在样品分析过程中的损失的程度，损失越少，回收率越高，如果作标液1PPM，就是1毫克/升，而作出标准数据为0.99毫克/升，就是说你的回收率是99%，这个与真实成分有密切的关系，说明方法的准确度。比如水中总无机氯含量测定，样品水中含有无机氯20mg/L，取100mL被测水样品，加入0.1mL浓度为10mg/mL的含无机氯标准样品，测定时忽略体积变化，如果测定出样品中无机氯为2.98mg/L，则认为回收率为99%。向左转|向右转
酶联免疫吸附（ELISA）实验是一种观察生物活性配体间反应活性的有效的观察手段，被广泛应用于疾病诊断和科学研究领域。Elisa试剂盒应用的范围很广，而且正在不断地扩大，原则上ELISA试剂盒可用于检测一切抗原、抗体及半抗原，可以直接定量测定体液中的可溶性抗原。Elisa试剂盒的灵敏度、特异性、精密度、稳定性等因素都会影响Elisa检测的结果，因此，在科学研究领域里，如何选择高质量的Elisa试剂盒产品成为了科研实验中的关键。
目前国内外Elisa试剂盒生产厂家很多，产品质量参差不齐。依据品牌网大数据平台数据支持，筛选出2016-2017年Elisa试剂盒十大品牌榜如下：
R&DSystems
R&DSystems经过近40年的发展，目前在英国、德国、法国和中国等地设有分公司。作为细胞因子、抗体和ELISA试剂盒等细胞生物学产品的顶尖供应商，R&DSystems在全球范围内为癌症、干细胞、免疫学、神经学和信号转导等领域的科研人员提供了品种齐全，质量可靠的科研产品，并因产品质量的可靠性和稳定性赢得了用户的广泛推崇和同行的赞誉是目前国内最受欢迎的Elisa进口试剂盒品牌。
Abcam
Abcam总公司位于英国剑桥，2014年1月成立艾博抗（上海）贸易有限公司负责中国区的销售。Abcam在中国设立的公司产品线包括一抗、二抗、免疫测定及细胞检测试剂盒、蛋白/多肽、激动剂/拮抗剂/活化剂/抑制剂及裂解物等。Abcam新推出的Elisa试剂盒使用重组单克隆抗体对，提供了其检测灵敏度。
BioVision公司是世界顶尖的细胞凋亡和信号传导产品生产商，产品包括Elisa试剂盒、荧光检测试剂盒、抗体及重组蛋白，几乎涵盖了细胞凋亡、细胞信号传导和细胞因子研究的各个方面。BioVision产品的价格虽然不菲，但其质量稳定可靠，在开发和研究细胞凋亡检测分析、代谢检测方面具有卓越的影响力，一直是研究人员在细胞检测领域的首选。
Raybiotech
Raybiotech公司主要涉及蛋白质芯片、抗体芯片、抗体、免疫试剂、Elisa试剂盒等近万种产品。RayBiotech生产超过2,600个经过ISO认证和严格测试的ELISA试剂盒，并得到了世界各国众多科学家的亲赖，许多客户对公司芯片技术和产品的质量都给予了肯定，客户发表的论文不乏Nature,NatureMedicine,Cell,PNAS，Lancet等世界顶级期刊物。
AssayPro
Assaypro是致力于为在学术界和制药/生物技术产业的研究人员提供高品质的免疫检测试剂盒。产品线包括AssayMaxELISA试剂盒，AssaySense活性测定试剂盒，抗体和其他相关试剂。AssayPro所有酶联免疫试剂盒和抗体产品都在美国密苏里州圣查尔斯ISO9001:2008和ISO13485:2003质量管理体系生产，凭借其在免疫科学领域独特的竞争优势，在国内有着比较广泛的市场群体。
PeproTechInc.创立于1988年，始终致力于为生命科学研究开发重组蛋白及相关产品，如今已成为能提供多种高质量细胞因子和重组蛋白，相应的单克隆、多克隆和生物素标记抗体，以及ELISA试剂盒的世界领导者。PeproTechInc.的产品涵盖免疫调节蛋白、白介素、集落刺激因子、生长因子、化学趋化因子、干扰素、脂联素、成纤维生长因子、转化生长因子、肿瘤坏死因子、神经营养素和防御素等诸多生命科学研究领域中重要的生物活性分子。
BioRike
BioRike博瑞克是目前国内非常优质的Elisa试剂盒厂家，近年来发展起来的名族品牌。BioRike产品线包含ELISA试剂盒、免疫组化试剂盒、抗体、快速基因克隆试剂等，其生产的Elisa试剂盒，灵敏度、精密度、准确度等综合性能指标处于行业领先水平，已广泛用于生命科学各领域科研机构，并获得广大科研人员好评，ELISA试剂盒自推出以来，已被大量权威杂志引用，在细胞因子、激素调控研究方面有着极强的竞争力。博瑞克试剂盒凭借其稳定的质量和亲民的价格成为国产明星品牌，在国内拥有一大批的忠实客户。
ExCellBio
依科赛生物目前主要有三大产品线：制药和生命科学研究用细胞培养系列产品、围绕基因检测的核酸产品线和免疫检测产品线。免疫检测产品线包括高品质的细胞因子ELISA检测试剂盒和抗体等，Elisa试剂盒在国内高校应用得比较广泛。
MultiSciences
联科生物与多家国际知名厂商达成了长期合作协议，包括Affymetrix（eBioscience）、Qiagen、Bio-Techne（RnD、Novus）、Roche等。在将国际上优质产品引入国内市场的同时，联科生物也在不断开发拥有自主知识产权的项目与产品，联科生物推出了两万多种抗体、辅助试剂以及成套Elisa试剂盒，这些产品主要应用于蛋白免疫印迹、流式细胞术等实验方法。联科生物的Elisa试剂盒自推出后，获得了市场的一致好评。
CUSABIO
华美生物主要经营抗体、蛋白、基因、科研试剂盒、诊断试剂原料产品。旗下的CUSABIO试剂盒，涵盖20个种属，数千种产品，涉及肿瘤、激素、自身免疫、心血管、代谢等十多个研究领域。华美生物的Elisa试剂盒价格相对偏高，对于国内科研经费不是特别充足的研究人员来说不是首选。
以上品牌都有各自的核心Elisa试剂盒产品，科研工作者可根据自身的研究领域的需求选择高质量的试剂盒品牌，以提升检测效率及结果有效性。
