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Q: Is your ViraDuctin™ AAV Transduction Kit compatible with all cell types?
A: Our ViraDuctin™ AAV Transduction kit works by affecting DNA synthesis, the rate limiting step of viral transduction, and is not cell type or AAV serotype specific. We have not tested it on all cell types; it should…
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Q: Will your AAV Purification kits work with any AAV serotype?
A: Our AAV purification kits use an affinity matrix to purify AAV-2 based on the structure of the AAV2 Cap protein; therefore, they only work for AAV-2 and AAV-DJ and are not suitable for purification of other AAV serotypes. Other…
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Q: Will the QuickTiter™ AAV Quantitation Kit work on all AAV serotypes?
A: This kit can be used on purified AAV of any serotype, including AAV samples that have undergone ultracentrifugation procedures. For crude, unpurified AAV it will only work on AAV serotype 2 and AAV-DJ.
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Q: How are these cells improved over normal 293 cells?
A: The HEK293 cell line is a permanent line established from primary embryonic human kidney transformed with human adenovirus E1 gene. 293AD is derived from the parental 293 cell line, but was specifically selected to have flattened…
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Q: What are the main differences between your two Adenovirus Purification Kits?
A: There are a couple of differences:
1. #VPK-099 uses a spin column while #VPK-5112 uses a purification resin.
2. #VPK-099 can purify one T75 flask or one 10cm dish per prep, while #VPK-5112 can handle up to…
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Q: Your ViraBind™ Adenovirus Purification Kit is listed as a 10 prep kit. Why does the kit only contain 5 filters?
A: The syringe-type filters found in this kit are designed to be used twice. Please see the product manual for the protocol to regenerate the filter for the second use, which will…
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Q: Which serotypes are your premade adenoviruses?
A: All of our Premade Recombinant Adenoviruses contain the Ad5 backbone with E1 and E3 deletions.
Q: What is the promoter for your premade adenoviruses?
A: All of our Premade Recombinant Adenoviruses use the CMV promoter.
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Q: How much adenovirus should be used for transduction?
A: The volume of supernatant required to infect cells will be different for every cell type and will be dependent on the MOI (multiplicity of infection) for that cell type. We recommend determining the MOI either by performing a…
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Q: What is the RAPAd® system?
A: The RAPAd® Adenoviral Expression System was developed by Dr. Beverly Davidson of University of Iowa. It uses an easy protocol that generates high titer virus in 2-3 weeks. Each RAPAd system contains a shuttle vector, a pacAD5 9.2-100 vector, a GFP control…
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Q: What cells can be infected with adenovirus?
A: Adenovirus is able to infect any cell type that expresses the CAR receptor, which includes most cell types. A literature search can be performed to determine if this receptor is expressed in a specific cell line or if other researchers have had…
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Q: Is there a positive control cell line that is easily infected by lentivirus?
A: We don’t have a control cell line, but we suggest using an easy to infect cell line such as 293T or our own 293LTV cell line. For ecotropic lentivirus we recommend NIH3T3 cells.
Q: How does your ViraDuctin™…
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Q: How are your 293LTV cells different from HEK293T cells?
A: The 293LTV cell line is a permanent cell line derived from HEK293T and was selected to have a high lentiviral yield. This cell line is fast growing and has a firm attachment to the culture plate, which can sometimes be problematic…
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Q: What is required to make lentivirus?
A: To make lentivirus, you need:
293T cells, such as our 293LTV cell line
A lentiviral expression vector to clone in a gene of interest
A lentiviral packaging system which includes vectors containing lentivirus structural proteins.
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Q: What is the difference between physical titer and infectious titer?
A: Viral titers are represented in two ways: either functional (infectious) titer measured in transduction units (TU/mL) or physical titer, measured in viral particles (VP/mL). Physical titer is a measurement of how much…
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Q: Can this kit be used with any lentivirus pseudotype?
A: Our Lentivirus-Associated p24 ELISA Kit uses two chemical polymers (Reagent A and Reagent B) that form a complex with the virus, independent of the envelope protein. These complexes are then pelleted, dissolved, and disrupted to…
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Q: How does this kit work?
A: Our ViraBind™ Lentivirus Concentration and Purification Kit (Column-based) uses a polymer technology that can concentrate the virus up to 500-fold with at least 60% recovery. Two chemical polymers are used that form a complex with virus at 37ºC. The complex is…
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Q: What is required to make retrovirus?
A: Retrovirus can be packaged using 293T or 293RTV cells. We also offer Platinum Packaging Cell Lines that can be used to package either amphotropic, ecotropic, or pantropic retrovirus. Here are the options for making retrovirus:
1. Transfect a Platinum…
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Q: How does this cell line differ from 293T cells?
A: The 293RTV cell line is a permanent cell line derived from HEK293 cells and was selected to have a high retroviral yield. This cell line is fast growing and has a firm attachment to the culture plate, which can sometimes be problematic with…
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Q: What is the difference between the three Platinum cell lines?
A: The Platinum cell lines are used to package retrovirus and they differ in the envelope protein that is expressed. Here is information about each of the Platinum cell lines:
1. Platinum-A cells express MMLV-Gag-Pol and an…
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Q: Which expression vector backbone should I use?
A: The retroviral expression vector will depend on the target cells being used. The pMCs vectors are used for ES and EC cells, the pMYs vectors are used with hematopoietic stem cells and progenitors, and the pMXs and pBABE vectors are suitable…
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Q: How does this kit work?
A: Our Retrovirus Quantitation Kit uses a polymer technology that pellets the virus from the media and uses a fluorescent dye to measure the retroviral RNA. The dye non-specifically binds to nucleic acid and is not virus-specific. However, any non-viral nucleic acids are…
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Q: How does your ViraDuctin™ Retrovirus Transduction Kit work?
A: ViraDuctin™ Retrovirus Transduction Kit is a reagent cocktail that uses technology developed at Harvard Medical School and can increase transduction efficiency 2-6 fold compared to polybrene. This product uses two chemical…
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Cell proliferation is an increase in the number of cells resulting from the normal, healthy process by which cells grow and divide. In this regard, cell proliferation can be a good indicator of general cell health. Cells that are subject to a variety of disease states may exhibit different rates…
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Q: Which detection method is more sensitive?
A: In our hands, the chemiluminescent and colorimetric formats display similar sensitivity. In general, luminescence usually offers better sensitivity and a broader range, and the chemiluminescent assay format may be more sensitive with certain…
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Q: Will this kit detect EGFP?
A: This kit will detect fluorescence from EGFP. The recombinant GFP standard used in the kit is also EGFP.
Q: What is the protocol for lysing cells?
A: Here is the protocol we recommend for lysing cells:
Wash cells once with cold PBS, then add cold 1X…