Adeno-1™ pShuttle(-) Cloning Kit
The Adeno-1™ Adenovirus Expression System provides reagents for efficient construction of recombinant adenoviral vectors with a CMV promoter. The principle of Adeno-1™ Adenovirus Expression System is to clone your gene of interest into the adenoviral genome by in vitro ligation (the same as in other subcloning projects). It is quicker, easier, and more reliable for recombinant adenovirus generation than in vivo homologus recombinantion used in previous approaches. Our subcloning shuttle vectors (pShuttle +/-) have more unique restriction enzyme sites than any other adenoviral cloning system that exists, offering you a variety of choices and simplifying your subcloning manipulation.
NOT FOR RESALE without prior written consent of ABM. This product is distributed for laboratory research only. * pricing for blank cloning vectors for commercial customers is 1.5x the listed price
|Caution||Not for diagnostic use.|
|Unit quantity||1 Kit|
- Selection-Drug Killing Curve
- Adenovirus Amplification/Transduction Protocol
|Can this kit purify all adenoviruses into the titer level at least 10^11 pfu/mL?|
Yes, if the insert gene in adenovirus is not toxic to cells. there are no difference in performance as regards to different kit and the difference is in the different amount of elution (final amount). However, high titer can be further concentrated with our buffer exchange kit.
|Would you have the titer data regarding adenoviruses that are lac g or cre recombinase gene-injected?|
We have in house experience with these two genes and they are not toxic, so higher titer adenovirus can be prepared.
|I have a question regarding preparation adenovirus stock for in vivo use. Adenoviral storage buffer in Viral stock buffer exchange kit (Cat.No.G130) is composed of 2.5% glycerol etc. which make it hard to inject in vivo in general. Accoriding to FAQs, 10mM Tris(pH8.0)plus 4% sucrose seems to be better formulation, but how can I remove 2.5% glyserol etc. from adenoviral storage buffer? How can buffer be exchanged from adenoviral storage buffer(Cat.No.G130) to 10mM Tris, 4% sucrose?|
In that case, you will have to go through the buffer exchange kit again using the sucrose buffer. A 3x20ml exchange will do the job.
|What are viral particle (VP), plaque formation unit (PFU), and infectious unit (IFU)?|
Viral particles (VPs) represent the total number of viral particles (infectious and infection-deficient combined). Due to variations in virus preparations, the ratio of infectious /non-infectious varies significantly and therefore, VP does not reflect the concentration of virus in a preparation. PFU (plaque forming unit) represents the number of infectious or live viruses. It reflects the concentration of infectious viruses in a preparation. IFU (infectious unit) is biologically equivalent to PFU. For most virus preps, the VP/PFU ratio is 20:1 to 50:1.
|How are virus titers determined?|
There are 3 commonly used protocols for determining adenovirus titer: (1) OD260 Assay, (2) Plaque Formation Assay, and (3) End-point Dilution Assay.
|How does TCID50 work?|
Please see the link before for the assay. TCID50 is the same as end dilution assay. http://www.abmgood.com/TechSupport/adeno-vec.php If the customer provide us the virus, we can provide its titer with the Cat#C008 service.
|Is this adenoviral gene transfer technique available for in vivo use? Is it OK to inject via tail vein?|
Yes, but higher titers up 1x10e12 pfu/ml have to be used. We do offer custom adenovirus amplification and purification.
|I'm wanting to overexpress my interest protein in mouse liver. Does this adenovirus transfer the gene specifically to the liver?|
Over 95% of adenoviral vectors will end up in liver if you inject in the blood stream.
|I'd like to perform a buffer exchange, but your Buffer Exchange column Cat# G130 seems to be discontinued. Can you offer me any recommendations for doing this?|
Unfortunately yes, our Buffer Exchange column G130 has now been discontinued due to a lack of supply for the filter component. In order to run your own buffer exchange you will require: 50K molecular weight cut off low protein binding membrane column (ensuring it will hold up to 15 ml) with a suggested buffer recipe as follows: 1 X PBS buffer, pH 7.2
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