Product Description
Advanced BioMatrix offers PhotoHA®, a methacrylated hyaluronic acid (HAMA) for photocrosslinkable hydrogels. These hydrogels provide native-like 3D HA gels with the unique attributes to be prepared at various concentrations and UV-crosslinked to provide various gel stiffness. The kit comes with 100 mg of lyophilized methacrylated hyaluronic acid and 100 mg of irgacure photoinitiator (365 nm UV photocrosslinking).
| Item | Catalog Number | Package Size | Storage Temperature |
| Methacrylated Hyaluronic Acid | #5212 | 100 mg | -20°C |
| Irgacure Photoinitiator | #5200 | 100 mg | 2-10°C |
Hyaluronic acid is the most abundant glycosaminoglycan in the body being an important component of several tissues throughout the body. While it is abundant in extracellular matrices, hyaluronic acid also contributes to tissue hydrodynamics, movement and proliferation of cells, and participates in a number of cell surface receptor interactions.
For the majority of cell types, it is recommended to add additional ECM proteins to the hyaluronic acid hydrogels. The proteins provide important cell binding sites.
Storage:
The product ships on frozen gel packs. Upon receipt, store the PhotoHA®at -20°C. Store the Irgacure at 2 to 10°C. The product and components are stable for a minimum of 1 year at receipt in powder form.
Once solubilized, the PhotoHA®can be stored at 2-10°C for 1 month. The photoinitiator can be stored for no more than 2 weeks once solubilized.
| Parameter, Testing, and Method | MethacrylatedHyaluronic Acid #5212 |
| Sterilization Method | Filtration |
| Sterility - USP modified | No growth |
| Form | Lyophilized Powder |
| Package Size | 100 mg |
| Storage Temperature | -20°C |
| Shelf Life | Minimum of 6 months from date of receipt |
| Shelf Life After Reconstitution | 1 Month |
| Degree of Methacrylation | 45-65% |
| Molecular Weight | 100-150 kDa |
NMR Analysis | Characteristic |
Hydrogel Young's Modulus E (Pa) | Characteristic |
Swelling Characteristics of PhotoHA®:
50 μL hydrogels fabricated in 4.7 mm diameter molds were imaged before and after incubation in phosphate buffered saline at 25°C for 24 hours. The diameter of hydrogels were quantified using ImageJ software. Statistical comparisons between groups (n=3) were performed via one-way ANOVA with post hoc testing and significance determined atp < 0.05.
Compressive Modulus of PhotoHA®:
Dynamic mechanical analysis (Q800, TA Instruments) was performed on 50 μL hydrogels fabricated in 4.7 mm diameter molds. Hydrogels were secured within a fluid cup via a 0.01 N pre-load and compressed to 30% strain at a rate of 0.5 N min-1. The Young’s modulus of each hydrogel was calculated as the slope of generated stress-strain curves between 10% and 20% strain. Statistical comparison between MeHA concentrations (n=3) was performed via Students t-test with two-tailed criteria and significance determined atp < 0.05.
Reaction Behaviorof PhotoHA®:
Rheological time sweeps (AR2000 stress controlled rheometer, TA Instruments; 0.5% strain, 1 Hz, 25°C) of MeHA crosslinking with exposure to UV light (=320-390 nm) and in the presence of 0.05 wt% Irgacure 2959 (I2959). After 1 minute, the macromer solution (i.e. MeHA and I2959) was exposed to UV light, resulting in the plateau of moduli before 5 minutes.
Directions for Use
Download the full PDF versionor continue reading below:
3D Hydrogel Preparation:
Note: Employ aseptic practices to maintain the sterility of the product throughout the preparation and handling of the collagen and other solutions.
Recommended concentrations are 5-30 mg/ml(0.5-3.0%).
Note: The following recommended instructions are for a 1% hyaluronic acid (HA) methacrylate solution. Adjustments to this protocol may be required for various concentrations.
- Add 10 ml of 1X phosphate buffer saline (PBS), water or cell culture media to the 100 mg of lyophilized methacrylated HA powder.
- Mix on a shaker table or rotator plate until fully solubilized (~30 to 60 minutes) at 2-10°C.Note: Solubilization times may vary depending on the desired concentration and volume of PBS, water or medium added,
- To solubilize the photoinitiator, add 1 mL of neat methanol to the vial of photoinitiator containing 100 mg of Irgacure and vortex. This makes a 10% solution.Note: The photoinitiator in solution has a shelf life of 2 weeks. Only dissolve required amount of photoinitiator required. Store remaining photoinitiator (powder or solution) at 2 to 10°C.
- Calculate the volume of the photoinitiator required by multiplying the total volume of HA methacrylate required by 0.01. For example, if you making 10 ml of HA methacrylate, the calculated volume of the photoinitiator to add is 100 µ
- Add the calculated volume of photoinitiator to the required volume of HA methacrylate solution and mix thoroughly.
- Add your cells to the HA methacrylate /photoinitiator solution.
- Dispense your HA methacrylate /photoinitiator/cell solution into the desired cultureware (i.e. 6-well plate, 48-well plate).
- For UV-crosslinking, place the hydrogel printed structure directly under a 365 nm UV light crosslinking source.Note: Longer exposure allows more crosslinking, though each cell type withstands different degrees of UV light and free radical exposure (generated by the photoinitiator) that mediates crosslinking
Product Q & A
Yes. There are quite a few publications citing various hyaluronidase protocols for digesting PhotoHA hydrogels. Here are a few:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460858/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4840832/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4986518/
The HA we usecomes frombiotechnological production, where the hyaluronic acid is extracted from the cell walls of the bacteria Streptococcus zooepidemicus.
Product Cell Assay
Human mesenchymal stem cells (20 x 106 / mL) were encapsulated in 50 μL hydrogels (~ 4.7 mm x 2 mm). Hydrogels (1 wt% PhotoHA®) were fabricated with 0.05 wt% Irgacure 2959 and exposure to 2 mW/cm2 light (320-390 nm) for 10 minutes. After 24 hours, encapsulated cells were stained with calcein AM and ethidium homodimer and subsequently imaged on a Leica SP5 confocal microscope (using FITC/TRITC sequential scans).
Product Applications
PhotoHA® Methacrylated Hyaluronic acid can be used to form hydrogels for ex-vivo engineering of autologous cartilage tissue[1] or as a mesenchymal stem cell carrier in cartilage repair[2].
Because the stiffness can be widely adjusted by altering concentration or UV-light exposure, methacrylated HA has been used to measure the effects of matrix stiffness on cell phenotype and function[3][4].
Methacrylated HA can be used for 3D bioprinting (extrusion[5], inkjet[5] and photolithographic[6]) to create structures that promote osteogenic differentiation of MSC’s[7].
The high tunability of hyaluronic acid methacrylate allows it to be mixed with, and reinforce other types of hydrogels (such as collagen, or gelatin methacrylate)[8].
References:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5717235/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627486/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5541838/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5447944/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615317/
https://www.ncbi.nlm.nih.gov/pubmed/21773726/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460858/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5748291/
Product References
References for PhotoHA®:
Khetan, Sudhir, and Owen Corey. "Maintenance of stem cell viability and differentiation potential following cryopreservation within 3-dimensional hyaluronic acid hydrogels."Cryobiology(2019).
Poldervaart, M. T.et al.3D bioprinting of methacrylated hyaluronic acid (MeHA) hydrogel with intrinsic osteogenicity.Plos One12,(2017).
Product Certificate of Analysis
Safety and Documentation
Satefy Data Sheet
Certificate of Origin
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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我用Promega公司的双荧光素酶检测试剂盒（E2920）检测到的firefly萤光素酶活性很低，只有4*10的3次方；海肾萤光素酶活性有10的6次方。
我用Ad293细胞做了转染，fireflyluciferase质粒：RanillaLuciferase质粒=0.1ug：0.025ug/一个孔（96孔板），共转染了二天，再进行双萤光素酶检测。
我想了解fireflyluciferase活性用promega的这个E2920-双萤光素酶试剂盒检测得到10的3次方，这种数值正常吗？
我做了3次重复实验，每次firefly萤光素酶活性很低，只有4*10的3次方；而海肾萤光素酶活性有10的6次方。
求指教？fireflyluciferase活性低？会是什么原因呢？
牛胰岛素,是一种多肽
在1965年9月17日我国完成了结晶牛胰岛素的全合成。经过严格鉴定，它的结构、生物活力、物理化学性质、结晶形状都和天然的牛胰岛素完全一样。这是世界上第一个人工合成的蛋白质，为人类认识生命、揭开生命奥秘迈出了可喜的一大步。这项成果获1982年中国自然科学一等奖。
1953年，英国人F. SangerSanger由于测定了牛胰岛素的一级结构而获得1958年诺贝尔化学奖。
1、 配制JC-1染色工作液：
取适量JC-1 Stain (200×)，按照每50μl JC-1 Stain (200×)加入8ml ddH2O的比例稀释JC-1，剧烈Vortex充分溶解并混匀JC-1。然后再加入2ml JC-1 Buffer(5×)，混匀后即为JC-1染色工作液。6孔板每孔所需JC-1染色工作液的量为1ml，其它培养器皿的JC-1染色工作液的用量以此类推。
2、 设置阳性对照：
推荐CCCP(10mM)加入到细胞培养液中处理细胞。随后按照下述方法装载JC-1，进行线粒体膜电位的检测。对于特定的细胞，CCCP的作用浓度和作用时间可能有所不同，需自行参考相关文献资料确定。
3、对于悬浮细胞：
a. 取1～6×105细胞，重悬于0.5ml细胞培养液中，细胞培养液中可以含血清和酚红。
b. 加入0.5ml JC-1染色工作液，颠倒数次混匀。细胞培养箱中37℃孵育。
c. 在孵育期间，按照每1ml JC-1 Buffer(5×)加入4ml蒸馏水的比例，配制适量的JC-1 Buffer(1×)，并放置于冰浴。
d. 37℃孵育结束后， 4℃ 600g离心3～4min，沉淀细胞。弃上清，注意尽量不要吸除细胞。
f. 再用JC-1 Buffer(1×)重悬后，用荧光显微镜或激光共聚焦显微镜观察，也可以用荧光分光光度计检测或流式细胞仪分析。
4、对于贴壁细胞：
注意：对于贴壁细胞，如果希望采用荧光分光光度计或流式细胞仪检测，应先收集细胞，重悬后参考悬浮细胞的检测方法。
a.吸除6孔板培养液，根据具体实验如有必要可以用PBS或其它适当溶液洗涤细胞一次，加入1ml细胞培养液。细胞培养液中可以含有血清和酚红。
b. 加入1ml JC-1染色工作液，充分混匀。细胞培养箱中37℃孵育。
c. 在孵育期间，按照每1ml JC-1 Buffer(5×)加入蒸馏水的比例，配制适量的JC-1 Buffer(1×)，并放置于冰浴。
d. 37℃孵育结束后， 吸除上清，用JC-1 Buffer(1×)洗涤2次。
e. 加入2ml细胞培养液，培养液中可以含有血清和酚红。
f. 荧光显微镜或激光共聚焦显微镜下观察。
5、对于纯化的线粒体：
a. 把配制好的JC-1染色工作液再用JC-1 Buffer(1×)稀释5倍。
b. 0.9ml 5倍稀释的JC-1染色工作液中加入0.1ml总蛋白量为10～100μg纯化的线粒体。
c. 用荧光分光光度计或荧光酶标仪检测：混匀后直接用荧光分光光度计进行时间扫描，激发波长为485nm，发射波长为590nm。如果使用荧光酶标仪，激发波长不能设置为485nm时，可以在475～520nm范围内设置激发波长。另外，也可以参考下面步骤6中的波长设置进行荧光检测。
d. 用荧光显微镜或激光共聚焦显微镜观察：方法同下面的步骤6。
6、荧光观测和结果分析：
检测JC-1单体时可以把激发光设置为490nm，发射光设置为530nm；检测JC-1聚合物时，可以把激发光设置为525nm，发射光设置为590nm。出现红色荧光说明线粒体膜电位比较正常，细胞的状态也比较正常。
注意事项：
1、 JC-1 Stain(200×)应完全溶解混匀后使用，但应避免反复冻融。必须先把JC-1 Stain(200×)用ddH2O充分溶解混匀后，才可加入JC-1 Buffer(1×)。不可先配制JC-1 Buffer(1×)再加入JC-1 Stain(200×)，否则导致JC-1很难充分溶解，严重影响后续的检测。
2、 对于6孔板中的样品，本试剂盒共可以检测100个样品；对于12孔中的样品，本试剂盒共可以检测200个样品。
3、 装载完JC-1后用JC-1 Buffer(1×)洗涤时，尽量使JC-1 Buffer(1×)保持4℃左右，此时的洗涤效果较好。
4、 勿把JC-1 Buffer(5×)全部配制成1×，因为操作过程中需直接使用JC-1 Buffer(5×)。
5、 如JC-1 Buffer(5×)中有沉淀，必须全部溶解后才能使用，为促进溶解可以在37℃加热。
6、 CCCP为线粒体电子传递链抑制剂，有一定毒性，请注意小心防护。
试剂盒在全球市场上的研发与销售呈快速上升趋势,2005年全球市场销售额超过200亿美元,且以15%左右的速度逐年增长.一方面是试剂盒的迅猛发展,而另一方面试剂盒市场良莠不齐的现象愈加明显,试剂盒的生产、销售及认证认可体制尚不完善,没有相应的标准或质量评价政策.且其灵敏度,稳定性及假阴/阳性控制尚不能满足检测需要,采用试剂盒进行检测的公信度受到质疑.
同时,食品安全领域是当前问题最多、最受关注的领域,这个领域的检测包括了物理、化学、微生物及分子生物学基础理论,无论是按检测原理、用途还是其它分类方式,涉及食品安全检测项目的试剂盒的品种是最全最多的.因此,从该领域着手从事评价制度的研究,便于获得基础性数据结果,并由此推广至动植物检疫及其它领域.
随着H7N9禽流感疫情的不断发散,国家流感中心已经多次发放人感染H7N9禽流感检测试剂,覆盖了全国31个流感网络实验室,并表示,诊断试剂的广泛发放是实现关口前移,控制疫情传播、蔓延的重要手段.而一旦H7N9监测关口继续前移,主动监测范围扩大,病毒检测试剂的需求量将进一步加大.
可采用H7N9亚型禽流感病毒RNA检测试剂盒（荧光PCR法）和H7N9禽流感病毒核酸检测试剂盒（PCR-荧光探针法）,定价分别为48人份/盒和48反应/盒,相比市场此前预期的100-200元之间的价格定位低了很多.在检验方法上,卫纪委提醒,前者需要配备全自动荧光PCR检测仪专用PCR扩增管和核酸分离试剂盒(硅胶膜吸附法)等必须设备及咽拭子样本,后者卫计委推荐采用达安基因生产的核酸提取试剂盒进行检验.
百奥森18分钟霉菌毒素检测试剂盒，有需要的可以了解下！多种规格型号可供选择
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实验室要开展支原体检测，方法是PCR法，先要采购试剂盒，用过的同学给推荐一下好用的品牌呗
暂无品牌问答
