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Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 100849 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 100849-AAV1 | Virus (100 µL at titer ≥ 7× 10¹² vg/mL)and Plasmid.More Information | Add to Cart |
This material is available to academics and nonprofits only.
Backbone
- Vector backbonepAAV(Search Vector Database)
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Copy numberUnknown
Gene/Insert
- Gene/Insert namejRCaMP1b
- Alt nameRCaMP1h variant 1519
- SpeciesSynthetic
- PromoterCAG
Cloning Information
- Cloning methodUnknown
- 5′ sequencing primerunknown (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- pAAV.CAG.Flex.NES-jRCaMP1b.WPRE.SV40 (p3850)_Q.gb
- Terms and Licenses
- UBMTA
- genOway Notice of RIghts
- Industry Terms
- Not Available to Industry
Depositor Comments
This plasmid was previously available as pAAV.CAG.Flex.NES-jRCaMP1b.WPRE.SV40 (p3850) from the Penn Vector Core. This plasmid was created as part of the GENIE project at Janelia Research Campus.
Information for AAV1 (Catalog # 100849-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV.CAG.Flex.NES-jRCaMP1b.WPRE.SV40 (#100849). In addition to the viral particles, you will also receive purified pAAV.CAG.Flex.NES-jRCaMP1b.WPRE.SV40 plasmid DNA.
CAG-driven, Cre-dependent NES-jRCaMP1b calcium sensor. 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, 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.
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分子生物学实验的染料主要涉及到核酸染料和蛋白质染料.核酸染料主要有EB(溴化乙锭,高致癌性),goldview,sybr green(实时定量PCR时常用染料).这些染料可以和核酸双链分子特异性结合发出强荧光而被检测到.蛋白质染料最常用的是考马斯亮蓝 R-250,硝酸银.其中硝酸银有时也用于核酸染色.
染料分为天然染料和人工染料两种。天然染料有胭脂虫红、地衣素、石蕊和苏木素等,它们多从植物体中提取得到,其成分复杂,有些至今还未搞清楚。目前主要采用人工染料,也称煤焦油染料,多从煤焦油中提取获得,是苯的衍生物。多数染料为带色的有机酸或碱类,难溶于水,而易溶于有机溶剂中。为使它们易溶于水,通常制成盐类。
染料可按其电离后染料离子所带电荷的性质,分为酸性染料、碱性染料、中性(复合)染料和单纯染料四大类。 标本干燥后即进行固定,固定的目的有三个:
1)杀死微生物,固定细胞结构。
2)保证菌体能更牢的粘附在载玻片上,防止标本被水冲洗掉。
3)改变染料对细胞的通透性,因为死的原生质比活的原生质易于染色。
是欢快活泼的光辉色彩,是暖色系中最温暖的色,它使人联想到金色的秋天,丰硕的果实;
是一种富足、快乐而幸福的颜色。让人感觉成一种稳重、含蓄又明快的温暖;
橙色也会带来一种甜甜的感觉。
希望同路人多多交流啊!
想请问一下,DAPI这个染料到底有没有膜通透性,我通过百度搜索查询关于DAPI染料的,基本上是说它能透过细胞膜对活细胞和死细胞均能染上蓝色;但是也有人说DAPI只可以透过死细胞膜,不能对活细胞进行染色,用以区分活死细胞,到底哪个是对的啊,蒙了!!!!!!
抗原抗体反应后,利用特殊仪器测定荧光强度而推算被测物浓度的检测方法
⑴荧光物质
1)荧光色素
许多物质都可产生荧光现象,但并非都可用作荧光色素。只有那些能产生明显的荧光并能作为染料使用的有机化合物才能称为免疫荧光色素或荧光染料。常用的荧光色素有:
⑴异硫氰酸荧光素(fluoresceinisothiocyanate,FITC)为黄色或橙黄色结晶粉末,易溶于水或酒精等溶剂。分子量为389.4,最大吸收光波长为490--495nm,最大发射光波长520--530nm,呈现明亮的黄绿色荧光,结构式如下:
有两种同分异结构,其中异构体Ⅰ型在效率、稳定性、与蛋白质结合能力等方面都更好,在冷暗干燥处可保存多年,是应用最广泛的荧光素。其主要优点是:①人眼对黄绿色较为敏感,②通常切片标本中的绿色荧光少于红色。
⑵四乙基罗丹明(rhodamine,RIB200)为橘红色粉末,不溶于水,易溶于酒精和丙酮。性质稳定,可长期保存。结构式如下:
最大吸收光波长为570nm,最大发射光波长为595~600nm,呈橘红色荧光。
⑶四甲基异硫氰酸罗丹明(tetramethylrhodamineisothiocyanate,TRITC)结构式如下:
最大吸引光波长为550nm,最大发射光波长为620nm,呈橙红色荧光。与FITC的翠绿色荧光对比鲜明,可配合用于双重标记或对比染色。其异硫氰基可与蛋白质结合,但荧光效率较低。
⑵其他荧光物质
1)酶作用后产生荧光的物质某些化合物本身无荧光效应,一旦经酶作用便形成具有强荧光的物质。例如4-甲基伞酮-β-D半乳糖苷受β-半乳糖苷酶的作用分解成4-甲基伞酮,后者可发出荧光,激发光波长为360nm,发射光波长为450nm。其他如碱性酸酶的底物4-甲基伞酮磷酸盐和辣根过氧化物酶的底物对羟基苯乙酸等。
2)镧系螯合物某些3价稀土镧系元素如铕(Eu3 )、铽(Tb3 )、铈(Ce3 )等的螯合物经激发后也可发射特征性的荧光,其中以Eu3 应用最广。Eu3
螯合物的激发光波长范围宽,发射光波长范围窄,荧光衰变时间长,最适合用于分辨荧光免疫测定。展开
荧光增白剂是一种荧光染料,也是一种复杂的有机化合物,其特性就是能产生蓝色荧光,可使肉眼看到的物质很白。
测试面膜是否含有荧光增白剂,打开面膜的包装,然后放到“ZF-C型三用紫外分析仪”下,使用紫外光照射面膜,看其是否会发出亮蓝色光,如果有,说明面膜上含有荧光剂。 如果没有紫外分析仪,可以用紫外手电筒,紫外验钞机/笔等。
含荧光剂的面膜在紫外光照射下,显现出非常大面积的亮蓝色。稍微接触面膜液体,就会粘附上荧光剂成分,拿了面膜的手,也有荧光。用清水冲洗手3遍后, 手上的荧光剂在紫外光照射下依然发出强烈的亮蓝色光,几乎没有洗掉。改用洗手液清洗3遍,照射后手上也发出蓝光,只是比清水的效果略好一些。用洗面奶清洗,与洗手液的效果差不多,都无法彻底清洗掉残留在手上的荧光...
荧光剂对人体的危害:
荧光剂不像一般化学成分那样容易被分解,而是在人体内蓄积,产生许多有害的作用,大大削减人体免疫力;荧光剂与伤口外的蛋白质结合,还会阻碍伤口的愈合;荧光剂能使人体细胞出现变异性倾向,其毒性累积在肝脏或其他重要**,会成为潜在的致癌因素。造成血液系统受损:化学物质容易污染人体血液,虽然血液具有一定的自净能力,微量的有害物质进入其中,会被稀释、分解、吸附和排出,但长期、大量的有毒物质倾注而入,必致其发生质的变化;进入血液循环,会破坏红细胞的细胞膜,引起溶血现象。