Lentivirus Promoter Blast™ kit

CAT.NOUNITPRICE
LV9504 x 1ml
$595.00

Specifications


Description

abm's Lentivirus Promoter Blast™ Kit is designed for easy monitoring and optimization of lentivirus transduction efficiency to help you achieve successful results in your stable cell line generation or engineering experiment. This kit includes 4 different GFP Lentiviruses (4 x 1ml at >1x106 IU/ml) with either the CMV, PGK, EF1α, or UBC promoter, enabling you to determine which promoter works best for your cell line. In addition, this kit can be used to calculate the optimal Multiplicity of Infection (MOI) for your transduction experiment - simply transduce your cells at various MOIs and monitor GFP signals!

Kit features:

  • Easy-to-use
  • Simplifies your transduction optimization process
  • Helps you draw accurate conclusions for which promoter is suitable for your cell line
  • Saves you time and resources

After the optimal infection condition and promoter is identified, the lentivirus with your target gene and under your chosen promoter may be ordered separately either in low or high titer format.

SKULV950
Unit quantity4 x 1ml
Titer>1x106 IU/ml
Note

Infection Protocol

1. Plate the target cells in a 24-well plate, 24 hours prior to viral infection at a density of 0.5×105 cells per well. Add 0.5 ml of complete optimal medium (with serum and antibiotics if required) and incubate the cells at 37°C with 5% CO2 overnight.

Note: It is possible to use other plate formats for transduction. In this case, the amount of cells should be adjusted depending on the growth area of the well/plate.

2. Thaw the lentivirus from –80°C at room temperature. We strongly suggest to make usable aliquots of lentivirus to avoid free-thaw since that is known to reduce the viral titer.Calculate the appropriate volume of virus needed to be diluted in media to achieve desired MOI. MOI= [(Product Titer (IU/ml) x Virus Volume (ml))/(Total Cell Number)]. Read more about Multiplicity of Infection (MOI) at our learning resources center.

3. Prepare a mixture of complete media with polybrene at a concentration of 8 μg/ml. Remove the growth media from the wells and replace with 0.5 ml of the polybrene-media-mix per well (add in ViralPlus Transduction Enhancer (Cat. No. G515) at 1:100 or at your own optimized dilution ratio).

4. Once an effective MOI has been determined for the target cells through preliminary test infections, use the appropriate volume of virus to infect your cells. Leave one well of uninfected cells as an additional standard control. Following the infection, incubate the cells at 37°C with 5% CO2 overnight.

5. The optimal MOI for the lentivirus can range from 1-100 depending on promoter and cell types and we suggest an MOI ranging from 0.5-5 as a starting point.

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References


2
  • Xia, et al. "High levels of protein expression using different mammalian CMV promoters in several cell lines" Protein expression and Purification. 45(1):115-124 (2006).
  • Wang, et al. "Impact of different promoters, promoter mutation, and an enhancer on recombinant protein expression in CHO cells" Scientific Reports : (2017).