Speedy Lentivirus Purification
Cat. No.
LV999
Unit
100 ml
Price
$162.00
| Cat. No. | LV999 |
| Name | Speedy Lentivirus Purification |
| Unit | 100 ml |
| Description |
Recombinant lentiviral vectors have been shown to be a powerful tool for stable gene transfer to both dividing and non-dividing cells in vitro and in vivo.Through years of experience with lentiviral vectors, ABM has developed its own proprietary pLenti-combo packaging mix and an efficient protocol for rapid production of recombinant lentiviral vectors with titers up to 107IU/ml. The quickest and easiest way to purify and concentrate lentiviral particles up to 100x.
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| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. LV999 |
| What is the mechanism of purification for the Speedy Lentivirus Purification? | |
|
The Speedy Lentivirus Purification is a positively charged matrix solution that binds to the negatively charged lentivirus, forming a precipitation that can be pelleted through centrifugation.
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| Can it also be used for Retrovirus purification? | |
|
No, because retroviruses are too sensitive to survive the required centrifugation step.
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| Will the lack of FBS and calcium affect the functionality of the speedy lentivirus purification? | |
|
No, the function will not be affected.
|
| What buffer should I use for resuspension? | |
|
The pelleted lentivirus can be resuspended in any appropriate buffer.
|
| Can I remove the matrix as much as possible from the resuspended lentivirus? | |
|
Yes it is possible but the lentivirus should go through a buffer exchange procedure. Our recommended method is loading the resuspended lentivirus onto a 20mL-50uL ultrafiltration unit (30,000 MWCO), add PBS, then wash out the matrix by multiple centrifuge spins and discard the flowthrough. Once the exchange has occurred, use the desired volume of solvent to resuspend the lentivirus from the top filter and aliquot into storage containers.
|
| What filter can I used for filter sterilization after the purification step? | |
|
We recommend a low protein binding filter with membrane pores of 0.2-0.45um. A few suggested membrane materials are PVDF, PES, and Supor (from Pall).
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| In the final solution what medium and FBS should I use to keep the concentrated viruses? | |
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When using our Speedy Lentivirus Purification (Cat#LV999), the choice of which medium and FBS to use to keep the concentrated viruses will vary based upon the intended downstream application of the lentivirus; i.e. depending on the complete media required for your cell line of interest. You can re-suspend the pellet in the complete media of the cell line which will be infected with this virus prep, to avoid dilution. For example, if 293T cells were the intended cell line, we would recommend using DMEM + 10% FBS. If in doubt, serum-free DMEM is typically a safe choice, however the required dilution will be a factor to consider.
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- Ikeuchi, W., Wakita, Y., Zhang, G., Li, C., Itakura, K., & Yamakawa, T. (2021). AT‐rich interaction domain 5A regulates the transcription of interleukin‐6 gene in prostate cancer cells. The Prostate, 82(1), 97–106. Portico. https://doi.org/10.1002/pros.24251
- Grimsdell, B., Saleem, A., Volpe, A., & Fruhwirth, G. O. (2023). Genetic Engineering of Therapeutic Cells with the Sodium Iodide Symporter (NIS) to Enable Noninvasive In Vivo Therapy Tracking. Positron Emission Tomography, 303–330. https://doi.org/10.1007/978-1-0716-3499-8_18
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