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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.
Q…
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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…
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Q: Will this kit detect EGFP?
A: The polyclonal antibodies used in the GFP ELISA kit (AKR-121) are against Jellyfish GFP (Aequorea Victoria) and will detect its variants EGFP, YFP, BFP and CFP, which are only point mutations of wild type GFP. The recombinant GFP standard used in the kit is…
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Q: How does the His-Tag Protein ELISA work?
A: Our His-Tag Protein ELISA (AKR-130) is a competitive ELISA that generates a reverse curve, with the lowest OD values obtained from high His concentrations. First, the sample is added to a plate coated with polyhistidine. The antibody is added…
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Q: What species is this kit compatible with?
A: Our Rapid Antibody Purification Kit is a protein A column based purification kit that can be used to purify IgG from human, mouse, rabbit, pig, dog or cat.
Q: Can I use this kit with more than 5 mL of sample?
A: The capacity of the…
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Q: Will the proteins retrieved with this kit be in their native forms?
A: The proteins retrieved using this kit will be renatured and properly folded, since only correctly folded proteins are able to bind to the glutathione beads.
Q: What is the optimal detergent ratio?
A: Every GST…
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Q: What GFP variants will this antibody recognize?
A: Our anti-GFP antibody is a mouse monoclonal raised against the N-terminal of GFP and will recognize native and denatured GFP and its variants, including EGFP, YFP, EYFP, and CFP.
Q: Does this antibody cross react with dsRed or mCherry…
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Q: Does this antibody cross react with EGFP?
A: Our anti-RFP antibody is a monoclonal antibody raised against the N-terminal peptide sequence of DsRed (VNGHEFEIEGEGEGR). Based on the peptide sequence alignment with EGFP, there is some degree of similarity and there may be some cross reactivity…
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Q: How were these cell lines made?
A: Our stable cell lines had GFP introduced by either lentivirus transduction or plasmid transfection, followed by selection.
Q: Can these cell lines be used in vivo?
A: Although we have not performed any in vivo experiments with these cells, we have many…
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Q: What is the recommended lysis buffer?
A: Cells and tissues can be lysed in RIPA buffer or with the following lysis buffer we have used: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM 2-glycerophosphate, 1 % Triton X-100 or 1 % Nonidet P-40, 1 mM EDTA, 1 mM EGTA, 1 mM Na3VO4 and Proteinase…
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Q: I am concerned about being able to detect low levels of active small GTPase in my sample using your Small GTPase Activation Assays. Do you have any suggestions to ensure I get good results?
A: There are two things you can do to improve your results with our Small GTPase Activation Assays…
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Q: Are the Small GTPase Assay Beads (Raf-RBD, PAK1-PBD and Rhotekin-RBD) species specific?
A: Sequence alignment of a specific small GTPase indicates that there is at most one or two amino acid variation between various species. Therefore our beads and also our primary antibodies may be used…
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Q: Are your Alkaline Phosphatase Kits dependent on species or cell type?
A: All of our Alkaline Phosphatase kits may be used for any cell type that expresses alkaline phosphatase such as osteoblast cells. The kits are not dependent on either cell type or species.
Q: I used your StemTAG…