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Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 104491 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 104491-AAV1 | 100 µL at titer ≥ 1×10¹³ vg/mLand Plasmid.More Information | Add to Cart | |||
AAV9 | 104491-AAV9 | 100 µL at titer ≥ 1×10¹³ vg/mLand Plasmid.More Information | Add to Cart | |||
AAV Retrograde | 104491-AAVrg | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV PHP.eB | 104491-PHPeB | 100 µL at titer ≥ 1×10¹³ vg/mLand Plasmid.More Information | Add to Cart |
This material is available to academics and nonprofits only.
Backbone
- Vector backboneAAV-Syn-FLEX(Search Vector Database)
- Backbone manufacturerScott Sternson
- Backbone sizew/o insert(bp)4894
- Total vector size (bp)6247
- Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature30°C
- Growth Strain(s)NEB Stable
- Copy numberLow Copy
Gene/Insert
- Gene/Insert namejGCaMP7s
- Alt nameGCaMP3-A52V K78H T302L R303P A317L D380Y T381R S383T R392G
- Alt nameGCaMP3 variant 1473
- SpeciesR. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish)
- Insert Size (bp)1353
- PromoterSynapsin
- Tag/ Fusion Protein
- T7 epitope, Xpress tag, 6xHis
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBsmBI(destroyed during cloning)
- 3′ cloning siteBsmBI(destroyed during cloning)
- 5′ sequencing primerACCACGCGAGGCGCGAGATAG (Common Sequencing Primers)
Resource Information
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- genOway Notice of RIghts
- Industry Terms
- Not Available to Industry
- Article Citing this Plasmid
- 1 Reference
Information for AAV1 (Catalog # 104491-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.
Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV9 (Catalog # 104491-AAV9)(Back to top)
Purpose
Ready-to-use AAV9 particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.
Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV9 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV9
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV Retrograde (Catalog # 104491-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.
Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons.These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 7×10¹² vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV retrograde (AAVrg)
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV PHP.eB (Catalog # 104491-PHPeB)(Back to top)
Purpose
Ready-to-use AAV PHP.eB particles produced from pGP-AAV-syn-FLEX-jGCaMP7s-WPRE (#104491). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7s-WPRE plasmid DNA.
Synapsin-driven, Cre-dependent, GCaMP7s calcium sensor.These AAV were produced with the PHPeB serotype, which permits efficient transduction of the central nervous system. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, PHP.eB cap gene pUCmini-iCAP-PHP.eB (plasmid #103005)
- BufferPBS + 0.001% Pluronic F-68
- SerotypePHPeB (plasmid #103005)
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Citation Information: When using the PHP.eB serotype in future publications, please acknowledge Viviana Gradinaru and cite Chan et al., Nat Neurosci, 20(8):1172-1179. Pubmed.ebiomall.com






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IgE的特点:IgE是血清浓度最低的免疫球蛋白 ,只有血清中IgG浓度的万分之一。IgE对热不稳定,是半衰期最短一的免疫球蛋白 ,只有2.8天,与细胞表面结合的IgE半衰期稍长,8~14天,IgE由变应原入侵部位(鼻咽、支气管、胃肠道)的黏膜固有层中的浆细胞合成。在各类免疫球蛋白中,IgE是合成率最低、分解率最高的。属于亲细胞抗体,过敏体质者的胎儿脐带血中IgE浓度可能升高,检测脐血中IgE浓度可用于评估胎儿过敏体质的可能性。
IgE检测方法:通常用ELISA方法检测总IgE。由于血清IgE浓度很低,一般酶免疫试验方法的敏感性不足以检出血清IgE,现在常规实验室检测血清IgE的试剂盒采用生物素——抗生物素蛋白 放大的ELISA。试剂盒中所含用于制定标准曲线的IgE标准品和检测结果的IgE浓度单位与其它免疫球蛋白 不同,不是用mg/L表示,而是用u/ml或ku/l表示。
IgE的正常值(参考范围):血清IgE水平在正常人群中呈偏态分布,即多数人为0或接近于0,IgE水平越高的人数越少。因此计算平均值时应计算几何平均值才能反映其真实情况,即用对数转换后其分布才能近似正态分布。
健康人群血清IgE水平与年龄关系较大,小儿和老年人的IgE水平低于成年人。新生儿血清中IgE水平很低,接近于零。随年龄增长,IgE水平也不断升高,5~7岁后接近正常人水平。按Pharmacia公司提供的参考范围,1个月以内<12KU/L,1岁<11KU/L,2~4岁<33KU/L,5岁以上至成人<85KU/L.
过敏性疾病患者的血清IgE水平可达2000~8000KU/L,当IgE水平高于2000KU/L时应考虑寄生虫感染.
有时血清总IgE水平检测结果为0或参考范围内低值,并不能排除过敏性疾病的可能,须结合临床表现和血清特异性IgE检测结果进行判断.
什么是特异性IgE检测(sIgE)?
通常所称的过敏原检测,并非真正检测血液样本中的过敏原分子,而是间接地检测其中针对某种过敏原的特异性IgE分子,特异性IgE检测实际上是检测过敏原特异性IgE,即检测样本中针对某种变应原的特异性IgE,从而间接地判断患者是否对某种过敏原过敏。
环境中常见的过敏原包括以下类别:
寄生虫和微生物:各种螨类(屋尘螨和粉尘螨等)、各种真菌(点青霉、烟曲霉、分枝孢霉、交连孢霉等)、蟑螂。
植物花粉:各种草花粉(豚草、葎草、蒿草)、各种树花粉(桑树、柏树、悬铃木、桦树、榆树、柳树、杨树等)。
动物皮毛:猫、狗、马、鸽子等动物的毛和皮屑。
过敏性疾病治疗的一场新革命
早在17世纪末,18世纪初,最早的显微镜被发明时,人们就已经知道人体内寄生着许许多的微生物,但是,那时的人们还没有意识到这些微生物在各种疾病的发生中扮演着怎样的角色。
直到19世纪末,一些科学家们提出了微生物致病学说,他们认为许多疾病是体内微生物造成的。这个革命性的理论提出后,医学家们开始努力识别那些能引起致死性疾病的微生物,如引起细菌性肺炎和结核病的微生物。医学家们还发明了许多药物试图杀灭这些可怕的致病微生物。
在这场人类与微生物进行的持久战争中,抗生素的发明可以说是人类取得伟大胜利的标志。从第二次世界大战末期开始,抗生素逐渐在世界范围内得到了广泛的应用,然而,从20世纪60年代开始,许多并非由病原微生物引发的慢性病或其它疾病的发病率急剧升高。过敏性哮喘就是一个典型的例子:没有任何证据证明感染某种病原微生物会引发哮喘,它也不具有传染性,但是在过去的40年间,其发病率却直线上升,成为儿童除呼吸道感染性疾病之后的发病率最高的常见病慢性病。
科学家们早在上世纪80年代末就开始注意这种奇怪的现象,有许多文献都提到了哮喘和过敏发病率上升的情况,但是直到最近,新的发现和新的理论才让人们意识到抗生素的滥用与这些疾病之间有着密切的关系,而且不是所有的微生物都会致病,许多事实表明,有些微生物有助于增强免疫力,可以预防或控制传统微生物致病学说无法解释的疾病。
益生菌对维持我们的健康至关重要。我们甚至认为可以把肠道内的一些微生物看做人体不可或缺的“器官”之一,无论它们所起的作用还是重要性,这些微生物完全可以和心脏、肺、肾脏等相提并论。这种器官最重要的功能 是使我们与日常生活中接触到的潜在有害微生物和平共处。如果这些微生物不能有效地发挥作用,人类也就无法保持健康,甚至无法生存。
对于儿童过敏性鼻炎、过敏性哮喘、儿童湿疹、儿童荨麻疹这些常见慢性过敏性疾病,非常困扰孩子的成长,值得我们提出的是,对于儿童过敏性疾病,除了药物治疗外,益生菌抗过敏免疫疗法,强调人体微生态平衡,认为过敏时应当将身体作为一个有机的整体看待,而不仅仅关注出现症状的那一部分,对于儿童过敏性哮喘发病率增高的病因中,医生们已经意识到“婴儿出生后过早使用抗生素,成为诱发过敏性哮喘发病的重要原因之一”,近十年,激素吸入剂已成为儿童过敏性咳嗽、过敏性哮喘的主要治疗手段,虽然吸入激素的治疗方法比起全身激素抗过敏副作用小,起效快,但是,我们仍然发现吸入激素的弊端,长期吸入激素的副作用以及过敏儿童治疗只局限于局部,得不到全面整个过敏反应系统的治疗,许多科学家不仅坚定地认同益生菌可以促进健康的观点,而且还发现了抗过敏益生菌菌种如唾液乳杆菌PM-A0006、格氏乳杆菌PM-A0005、约氏乳杆菌PM-A0009能有效平衡过敏免疫反应从而影响整个过敏性疾病的治疗进程。
由于这些安全、自然的抗过敏益生菌组合物康敏元抗过敏益生菌能够帮助我们应对过敏这种常见而又难以对付的过敏性咳嗽,过敏性哮喘等疾病。
呼吸道过敏症不完全是由过敏原引起的,真正的罪魁祸首是免疫系统本身。过敏体质患者的免疫系统不能正确识别花粉或真菌,它把这些无害物质识别成有害的微生物,进而发生了不应有的免疫应答:与哮喘发作一样,这些症状严重时可能会引发致命的后果。
抗过敏益生菌和过敏免疫系统的相关性研究
近年来,抗过敏益生菌调节免疫功能,平衡因TH2过度反应而引起的过敏反应这一机理在过敏性疾病免疫学领域产生了一个新理论,为这场关于抗过敏益生菌的革命铺平了道路。
康敏元抗过敏益生菌抗特异性IgE的非药物抗过敏免疫疗法
适合反复过敏或对多种物质过敏者以及有过敏体质者,对过敏性鼻炎,过敏性哮喘,过敏性咳嗽,荨麻疹,湿疹,异位性皮炎等过敏症状通过非药物抗过敏免疫疗法康敏元加强型益生菌对于湿疹(奶癣)的免疫调节改善一般是1-3个月,对于过敏性鼻炎过敏性咳嗽和过敏性哮喘的改善一般是40-60天会有自觉性过敏症状明显减轻的改善。
已有研究证实一些特定的益生菌如唾液乳杆菌和格氏乳杆菌经过完整临床测试:可定殖肠道刺激肠道内的免疫细胞进行免疫调节,对于过敏体质的调整发挥最佳功效。“益生菌调整过敏体质”早已在免疫学及医学界对导致过敏发生的根源已达成共识,根据基因的功能性分析,通过国际最新菌株筛选平台(SINT),运用基因芯片进行高效筛选及体外基因重组,找到了具有抗过敏基因的益生菌菌株,研究证实,唾液乳杆菌LS,格氏乳杆菌,约氏乳杆菌,副干酪乳杆菌、罗伊氏乳杆菌都具有显著的抗过敏功能,其中唾液乳杆菌是研究最多的抗过敏的乳酸菌分离菌株,唾液乳杆菌与格氏乳杆菌,约氏乳杆菌,副干酪乳杆菌等组成五种菌体复方抗过敏菌株群康敏元益生菌,明显缩小了抗过敏功能个体差异化,相比单一抗过敏菌株免疫调节抗过敏能力更加全面。
叶酸和生物素都属于B族维生素,
对维持机体的生理功能有重要作用。
求速度考试,简答
现配现用
1) 物素N—羟基丁二酰亚胺酯(biotinyl-N-hydroxy-succinnimideBNHS)制备:
取物素1g混悬于12 mlN,N-二甲基甲酰胺(DMF)加0.6g N-羟基丁二酰亚胺(HOSU)0. 8 g二环基碳二亚胺(DCC)置于密闭容器内室温磁力搅拌作用夜滤滤液经旋转蒸干加10mL乙醚洗涤继加200 ml异丙醇使其重结晶获白色粉末状晶体
BNHS酯键-C=0基团与蛋白质赖氨酸氨基结合使物素标记蛋白含赖氨酸残基越或蛋白质等电点pI 6.0其标记效越BNHS适用于标记抗体及性偏碱性抗原

