
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 107787 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV8 | 107787-AAV8 | Virus (100 µL at titer ≥ 1×10¹³ 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 nameCre
- PromoterTBG
Cloning Information
- Cloning methodUnknown
- 5′ sequencing primerunknown (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- pENN.AAV.TBG.PI.Cre.rBG(p1091).gb
- Terms and Licenses
- UBMTA
- Industry Terms
- Not Available to Industry
Depositor Comments
AV-8-PV1091 From University of Pennsylvania Vector Core
Information for AAV8 (Catalog # 107787-AAV8)(Back to top)
Purpose
Ready-to-use AAV8 particles produced from AAV.TBG.PI.Cre.rBG (#107787). In addition to the viral particles, you will also receive purified AAV.TBG.PI.Cre.rBG plasmid DNA.
TBG-driven expression of Cre. 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, AAV8 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV8
- 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.
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我要做的两个目的蛋白,分子量一个是140的,另一个是36,内参用的GAPDH。这几天刚刚要做小鼠组织的蛋白,就把新提取的组织蛋白和之前做过的细胞蛋白一起上了,细胞和组织的各上了3个孔。
电泳转膜孵育等条件都是以前做成熟的
之后显影,细胞蛋白的,两个目的和内参都出得挺好;但是小鼠的组织蛋白只有分子量140的那个显出条带了,36的那个显出来的像一片水渍一样,斑片状的黑,内参有时也这样、有时干脆什么都不显。不知道这是什么原因...按说细胞同样的蛋白做出来,WB的各个步骤应该不会有太大问题的。
有点怀疑蛋白提取的不好,我提组织蛋白的的操作是在冰上进行的,裂解液也加有PMSF,用BCA法测得的蛋白浓度大概也有3~5mg/ml。
在这里求助各位高手~望给我指点一下~不胜感激啊!
去垢剂作为添加剂生长蛋白质晶体是一项很有用的技术。非离子和两性离子结晶剂在膜蛋白结晶中已经得到很好应用,目前已经变成常规的实验手段。去垢剂在一些可溶蛋白中也起到了一定的作用,提高了晶体的质量和结果的可重复性 ,促进了单晶的生长, 但具体的影响机制尚不明确。其可能原因是去垢剂的引入使结晶液滴中的蛋白分子更具有亲水性,使溶液中同质蛋白质分子增加。向左转|向右转
如要膜蛋白保持原来的结构应采用:非离子型去垢剂.
表面活性剂:通常由一个极性头和一个非极性尾组成,用于去除难溶于水的污渍
漂白剂:通常含次氯酸盐成分,原理为其氧化性
稀盐酸:用于去除碱性的污渍,如马桶中的结块
碳酸钠溶液:用于去除油渍,其原理为酯在碱溶液中水解,参见皂化反应

