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GoldBio/SHH, Human/25 μg/6110-19-25

GoldBio/SHH, Human/25 μg/6110-19-25
  • GoldBio/SHH, Human/25 μg/6110-19-25
商品介绍

GoldBio growth factors are manufactured for RESEARCH USE ONLY and cannot be sold for human consumption!

Recombinant Human Sonic HedgeHogrHuShhHhg2

Members of the Hedgehog (Hh) family are highly conserved proteins that are widely represented throughout the animal kingdom. The three known mammalian Hh proteins, Sonic (Shh), Desert (Dhh) and Indian (Ihh), are structurally related and share a high degree of amino acid sequence identity. Each Hh protein has a unique expression pattern and distinct biological role within its respective region of secretion.All Hh proteins use the same signaling pathway and can substitute for each other in experimental systems. Shh has been found to have a critical role in development, acting as a morphogen in the patterning of limbs, brain structure, spinal cord, thalamus and the teeth.Shh has been shown to attract commissural axons in the developing spinal cord by attracting or repelling retinal ganglion cell (RGC) axons depending on concentration.

Formulation: Sterile filtered whitelyophilized powder. Purified and tested for use in cell culture.

Sequence: IVIGPGRGFGKRRHPKKLTP LAYKQFIPNV AEKTLGASGR YEGKISRNSE RFKELTPNYN PDIIFKDEEN TGADRLMTQRCKDKLNALAI SVMNQWPGVK LRVTEGWDED GHHSEESLHY EGRAVDITTS DRDRSKYGML ARLAVEAGFDWVYYESKAHI HCSVKAENSV AAKSGG

MW: ~20.0 kDa (176 amino acid)

Purity: >98 % bySDS-PAGE and HPLC analyses.

Biological Activity: Fully biologically active when compared to standard. The ED50 as determined by inducing alkaline phosphatase production of murine C3H/10T1/2 cells is less than1.0 μg/ml, corresponding to a specific activity of >1.0 × 103 IU/mg.

Accession Number: Q15465

HGNC ID: 10848

Source: Escherichia coli

Storage/Handling: This lyophilizedpreparation is stable at 2-8°C, but should be kept at -20°C for long termstorage. The reconstituted sample can be apportioned into working aliquots andstored at -80°C for up to 6 months.Avoid repeated freeze/thaw cycles.

Reconstitution: The sample shouldbe briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitutein a siliconized tube using PBS that contains a 0.1% BSA to a concentration of0.1-1.0 mg/mL. Reconstituted solutions are stable for up to one week at 2-8°C.Stock solutions should be aliquoted and stored at -80°C. Further dilutionsshould be made in appropriate buffered solutions.

Additional resources for this product may be found at:

OMIM

NOT FOR HUMAN USE!

品牌介绍

About GoldBio:

GoldBio Research

Gold Biotechnology shares a vision with every researcher: Each facet of life science research will introduce us to a future of impactful discovery and learning, benefiting the entire world. And our vision will materialize sooner if a researcher’s passion is not sullied by worry over the quality of their reagents, especially the cost for quality. This is why we’re here! We believe in a future of change and prosperity, and we want to help you get there by being a resource for high-quality products at exceptional prices. By providing our customers with only the best chemicals possible, tested in GoldBio labs to assure our own quality specifications are met, you can be sure your research will go off without a hitch.

Mission Statement:

"At GoldBio, we strive to exceed the expectations of our customers, vendors and strategic partners through quality of products, excellent service and attention to detail. Our sustainable business model and relationships with others in the supply chain allow us to provide researchers and scientists with uniquely low prices, while maintaining the highest quality in the industry. We stand behind all our products to the fullest extent with the hope that they will be used in discoveries that benefit all."

Our Philosophy:

Provide the highest quality product
Maintain low costs to pass along to the researcher
Be an advocate for the industry
Support our customers as any loyal business partner
Be an ally to businesses in the industry
Put forth products with hope
Look towards the future
We look toward the future with hope as we send out our products with the intent that they will be used in great discovery. We are a small company dedicated to the success of those whom we supply. So we provide high-quality low cost materials to scientists and researchers. We give our customers the best so they can discover, invent, and innovate. Because that is what it is about for us...The Discovery.

Commitment to Quality:

We are dedicated to providing our customers with the highest-quality products available. We test our products from our network of trusted global suppliers to assure they meet our quality specifications. We stand behind all of our products knowing that our reputation lies on the success of your experiments.

History:

Dr. Paul Gold

During the 1980s, Dr. Paul Gold stepped out from behind the bench believing researchers needed a low-cost, high-quality alternative for chemicals and reagents. Recognizing his knack for business, he began brewing the idea for a company during a time when startup incubators were nearly unheard of, and the Internet, social media and crowdfunding didn’t exist. Armed with a supportive wife, generous family members and helpful people within his network, Dr. Gold established GoldBio in 1986.

The original main office was situated in the former St. Louis Technology Center (now the location for the St. Louis Science Center), while the first lab space was located at St. Louis University. The ambitious startup began its journey selling three products: IPTG, X-Gal and X-Gluc.

Now, after almost 30 years, GoldBio has evolved significantly, expanding its catalog from three products to more than 3000 products. Throughout its history, GoldBio’s success has relied on its commitment to the success of those whom we supply. At GoldBio, we believe that putting our quality products in capable hands leads to discoveries that impact the entire world。

 


苏州蚂蚁淘生物科技有限公司是一家创新型高科技生物公司,以常规生物学实验外包与生物学试剂为基础,放眼于生命科学领域的前沿技术服务,公司技术骨干有着多年的生物学科研背景,毕业于国内外名牌大学,并且有着丰富的项目经验,了解国内外科研行情,我公司力求为每一个客户提专业、针对性的实验方案。我公司以技术服务为支点,和国内外多家生物试剂公司有着密切的业务往来.公司特色技术服务包括:抗体药物研发、噬菌体抗体库、蛋白表达与传化、抗体制备、高通量测序、组学研究、生物信息学分析与方法建立、基础分子生物学技术外包、化学中间体合成、原料药物研发、论文编写等。我公司秉承”诚信做人,认真做事,服务至上“,希望用我们的专业知识和项目经验为国内科研客户解决科研问题,祝您科研之路畅通无阻!


背景:活体成像技术(optical in vivo imaging)目前主要采用生物发光(bioluminescence)与荧光(fluorescence)两种技术,生物发光法是基于荧光素酶能催化底物(D-Luciferin或Coelenterazine)化学发光的原理,将体外能稳定表达荧光素酶的细胞株植入动物体内,与后期注射入体内的底物发生反应利用光学系统检测光强度,间接反映出细胞数量的变化或细胞的定位。这项技术已被广泛应用于多个领域,最常用的有肿瘤或疾病动物模型的建立,并可用于病毒学研究、siRNA研究、干细胞研究、蛋白质相互作用研究等。
原理:常见的荧光素酶有两种,分别是萤火虫荧光素酶(firefly luciferase,编码基因是luc)和海肾荧光素酶(Renilla luciferase,编码基因是Rluc),前者的底物是D-Luciferin,后者的底物是Coelenterazine。它们共同的作用原理是在ATP和荧光素酶的催化作用下,底物被氧化发光(不同底物光的颜色和波长不同),当底物过量时,产生的光量子数与荧光素酶的浓度呈正相关性。
产品:化学发光底物1:D-Luciferin美国Goldbio公司提供几种常用类型的D-Luciferin,每批试剂都经过严格的质量验证 (经9次独立的HPLC和FTIR标准法监测)和发光效率检测,确保客户每次试验的高重复性和成功性。所有产品都可用于体内成像分析和体外化学发光检测实验。Goldbio的D-Luciferin是合成的萤火虫荧光素,结构与promega公司提供的甲虫荧光素一致,是水溶性的萤火虫荧光素酶底物。DLuciferin, Sodium Salt/ D荧光素钠盐分子式:NaC11H7N2O3S2·H2O。 
分子量:320.32 g/mol     纯度:高级纯(99.7%)
应用:
1)体外化学发光分析(in vitro);
2)活体成像实验(in vivo);
3)高灵敏度ATP分析;
步骤:
Protocol 1In Vitro Bioluminescent Assays / 体外生物发光检测
1)用33.3 mL蒸馏水溶解1.0 g D-荧光素钠盐,配制成100 mM的储存液(200×,浓度30mg/ml)。混匀后立即使用或分装后-20℃冻存。
2)用组织培养基1∶200稀释储存液,配置工作液(终浓度150μg/mL)。
3)去除培养细胞的培养基。
4)待图像分析前,向细胞内添加1×荧光素工作液,然后进行图像分析。
Protocol 2:In vivo analysis / 活体成像分析
1)用无菌的PBS(w/o Mg2+、Ca2+)配制D-荧光素钠盐工作液(15mg/mL),0.2 um滤膜过滤除菌。一旦使用,保持冰冷且避光。
2)注射量取决于注射方式,具体如下:
注射方式剂量静脉注射(25-27 gauge针头)按10 uL/g体重浓度,加入相应体积的15 mg/mL荧光素工作液 腹腔注射(25-27 gauge针头)按10 uL/g体重浓度,加入相应体积的15 mg/mL荧光素工作液肌肉注射(27 gauge针头)50 uL,浓度为1–2 mg/mL荧光素工作液鼻内注射(pipette)50 uL,浓度为3 mg/mL荧光素工作液3)注射入体内5-10分钟后,进行成像分析。
(2)D-Luciferin, Potassium Salt/D-荧光素钾盐;
分子式:NaC11H7N2O3S2·H2O    分子量:320.32 g/mol   纯度:高级纯(99.7%)
应用:
1)活细胞、组织或生物体内luc标记基因和荧光素酶-融合基因体内/体外表达的成像分析;
2)广泛用于报告基因分析,免疫分析和ATP荧光卫生监测分析;
Protocol 1:In Vitro Bioluminescent Assays / 体外生物发光检测
1)配制成100 mM的储存液(200×,浓度30mg/ml)。混匀后立即使用或分装后-20℃冻存。
2)用预热好的组织培养基1∶200稀释储存液,配制工作液(终浓度150μg/mL)。
3)去除培养细胞的培养基直至无残留。
4)图像分析前立即向细胞内添加1×荧光素工作液,然后进行图像分析(或者细胞放在37℃短时间孵育后检测可增强信号)。
Protocol 2:In vivo analysis in mice / 小鼠活体成像分析
1)用无菌的DPBS(w/o Mg2+、Ca2+)配制D-荧光素钾盐溶液(15mg/mL),0.2 um滤膜过滤除菌。一旦使用,保持冰冷且避光。
2)按10 μL/g体重的浓度向每支小鼠注射相应量的15mg/mL荧光素溶液;
3)腹腔注射10-15 min后进行成像分析(对每只动物模型做荧光素动力学研究以确定峰值信号获取时间
(3)D-Luciferin Firefly, free acid/D-萤火虫荧光素
分子式:C11H8N2O3S2  分子量:280.330 g/mol  外观:白色至浅黄色粉末
溶解:0.25%浓度溶解于甲醇,形成近乎无色至浅黄色的清夜
保存:-15°C 避光
应用:
1)活细胞、组织或生物体内luc标记基因和荧光素酶-融合基因体内/体外表达的成像分析;
2) 广泛用于报告基因分析,免疫分析和ATP荧光卫生监测分析;
使用GBT公司荧光素(Luciferin)发表的文献:
1. Barth et al. Molecular Therapy. 2008, 16. 957-964.
2. Mahller et al. Cancer Research. 2008, 68 (4). 1170-1179.
3. Marques et al. Journal of Neurovirology. 2008, 14 (3). 229-238.
4. Stish et al. J. Neurooncol. 2008, 61 (1). 51-61.
5. Zhang et al. Cancer Research. 2007, 67. 9389-9397. 6. Baia, G. et al, Brain Pathology, 2007, 18 (2), 172-179.
7. Barth, A. et al, Molecular Therapy, 2008, 16 (5), 957-964.
8. Su Y. et al. Clinical and Experimental Metastasis, 2009
化学发光底物2:Coelenterazine
美国Goldbio提供三种coelenterazine类似物,分别是Coelenterazine,Coelenterazine h,Coelenterazine 400a。虽然三种都可用于相同实验,但是得到的结果有差异。
Coelenterazine:普遍认为是天然的腔肠荧光素,是海肾荧光素酶(Rluc)的作用底物,具有许多用途。可用于检测活细胞中钙离子浓度,基因报告分析,BRET(生物发光共振能量转移)研究,ELISA,HTS,组织或细胞中超氧阴离子和过亚硝酸盐的化学发光检测。
Coelenterazine h: 与Renilla荧光素酶或Gaussia荧光素酶使用具有一定的局限性。体外研究中比其他类似物更适合与钙离子激活发光蛋白共用。据报道,Coelenterazine h的发光强度高于天然腔肠荧光素10-20倍,因此是用来检测钙离子浓度微小变化的有力工具。
Coelenterazine 400a: 是BRET研究中Rluc的首选底物,此腔肠素衍生物在390 nm和400nm之间发蓝色光,对GFP acceptor的光发射干扰最小。
(1)Coelenterazine(天然腔肠荧光素)
分子式:C26H21N3O3    分子量: 423.48 g/mol   纯度:95%  保存: -15°C避光,保存在惰性气体下
Protocol:发光检测实验
1)使用管光度计室温下定量检测发光情况,每个样品做三个平行;
2)加入底物10s内立即检测,“integration time”为1 s;
3)对于细菌表达Gluc系统,甲醇溶解底物腔肠素制备10 mM储存液,终浓度为10μmol/L。储存液-20 °C保存,工作液制备过程需避光并置于冰上。
(2)Coelenterazine 400 a/腔肠素 400a
分子式:C26H21N3O   分子量:: 391.48 g/mol     纯度: 95%
储存液制备:
1)开封前将瓶子放到室温温育,以避免产品发生任何的冷凝;
2) 用125 μL无水乙醇溶解50 μg Coelenterazine 400 a,最终浓度是0.4μg/μL. ,轻轻混匀:避免用DMSO或其他溶剂来溶解腔肠素400a,因为试剂可能被氧化)
3)分装后避光保存在-20°C,保持干燥;
(3)Coelenterazine h/ 腔肠素h(from Caliper LifeScience Inc.)
RediJect Coelenterazine h为即用的注射型生物发光底物,特别设计用于体内成像研究。通过最小化产品成分的变化,最小化冻融次数和严格控制不同批次的产品,使用Caliper公司的Coelenterazine h能确保客户得到高重复性的结果。
优  点:
1)预制配方,不同批次间严格控制特别设计用于体内研究;
2) 即用型产品通过最小化预成像准备步骤,减少时间和精力;
3) 每瓶可用于5只动物的成像研究;(10 瓶/kit)
4)经IVIS成像系统验证,控制体内成像质量;
颜 色:黄色溶液(溶于丙二醇/柠檬酸盐)
浓 度:150 μg/mL (10瓶无菌溶液,每瓶含1.2 mL浓度为 150 μg/mL的 Coelenterazine h )
保 存: Store at ≤ -70 °C。
使用及注射剂量:
开始实验前,37 °C 水浴锅解冻,避免重复冻融。静脉注射(i.v.)或腹腔注射(i.p.)。
1)静脉注射:推荐使用剂量15 μg/mouse(100 μL/mouse),用1 ml注射器直接从瓶内吸取溶液,然后用25 gauge或更高的针头注射入小鼠体内。尽快成像检测以获得峰值信号。
2)腹腔注射:推荐剂量是30 μg/mouse (200 μL/mouse)。

品名 目录号 价格
D-Luciferin,Sodium Salt LUCNA-100 1400
D-Luciferin,Sodium Salt LUCNA-500 4000
D-Luciferin,Sodium Salt LUCNA-1G 6000
D-Luciferin,Potassium Sal LUCK-100 1400
D-Luciferin,Potassium Sal LUCK-500 3000
D-Luciferin,Potassium Sal LUCK-1G 49000
D-Luciferin Firefly,free acid L-123-100 1400
D-Luciferin Firefly,free acid L-123-500 3000
D-Luciferin Firefly,free acid L-123-1G 49000
Coelenterazine CZ1 1900
Coelenterazine CZ5 3800
Coelenterazine CZ10 4600
Coelenterazine 400a C-320-1 2200
Coelenterazine 400a C-320-5 6400
Coelenterazine 400a C-320-10 1200

 
 
   
 


 
 
 
 
 
 
 
 
 
 
 
 


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