1

Welcome to ATCGbio Life Technology Inc.

Advancing Life Sciences Technology

pLenti™ Lentivirus Production Kit (LT1001)

Availability: In stock

USD$400.00
OR

Quick Overview

 Catalog Number          Size                 Price (USD)


      LT1001                  3×preparations*              400.00

* 3 preparations mean that the amount of reagents are enough to make lentivirus in three 6-well plates. One well of 6-well plate will produce 4ml lentivirus at titer up to 0.9 x 108 TU/ml. 4ml lentivirus are enough for experiments in two 6-well plates at highest amount of lentivirus added (300μl lentivurs per well). At this amount of lentivirus, the primary cells or differentiated cells will be infected more than 70% (the most of those cells will be 90% infected). Some primary cells or differentiated cells just require 50-100μl lentivirus per well to get suffecient infection (>70%).

Download:   USER MANUAL PDF  MSDS PDF
Highlight the Kit Features
1. Small scale (6 well plate format), high titer (up to 0.9 x 108 TU/ml) virus production system, which in turn to save physical and financial expenditures. 
2. The provided production medium does not contain chemicals used to add lentivirus production such as sodium butyrate, caffeine, or chloroquine. So, lentivirus in this medium can be directly used for most of applications. 
3. Reagents for concentration, purification and long-term storage are included. 
4. The kit produces high quality virus, meaning that higher percentage of virus is functional, resulting in higher transduction rate. High quality of virus is more important than virus number (assessed by p24 antigen, RT-PCR). 
5. Purified virus can be injectable locally in experimental animals.
6. Compatible with both 2nd. and 3rd. generation of virus constructs.
7. The kit includes GFP expressing lentivirus plasmid.

pLenti™ Lentivirus Production Kit (LT1001)

Details

This kit is particularly suitable for users using primary cells. In many occasions, it replaces time-consuming adenovirus system because of short preparation time and ease of use. User provides lentivirus vector plasmid (2nd. or 3rd.  generation), and in 48 hours lentivirus is ready. The kit is designed for 6-well plate and can produce 4 ml lentivirus per well with 0.5~9 x107 TU/ml titer. This amount is sufficient to infect the cells growing 100 mm. Although freshly prepared virus showed the highest titer, it still can be stored up to 4 days at 4 °C and 2 weeks at -20°C. For long-term storage, it is recommended to use the provided storage solution after x1500 g centrifugation.

With our pLSLIC kits, user can create gene of interest expressing lentivirus vector plasmid in just one day. We also provide pre-made and custom-made shRNA lentivirus plasmids.

The crude virus can be used directly to infect target cells if the medium (DMEM base) is compatible and the cells are ready. In case that medium is not compatible or if users want to store the virus longer time (crude virus keep sufficient titer in 4 days at 4°C, in 2 weeks at -20°C), the kit also provides purification reagents which allows to produce concentrated purified virus with high stability in -80oC storage. With this unique configuration and optimal production reagents, the user can use lentivirus as flexible as adenovirus in both primary and transformed cell lines.

Fig.A and Fig.B (magnification 100x): GFP expressing lentivirus was created with two different medium

    Conventional DMEM+10% FBS                              Our Lentivirus Production Medium + 2% FBS

 Lentivirus-infect293T-cell(ordernary-medium-cell-culture) Fig.A  

       Lentivirus-infect293T-cell(our-Production-Medium-culture) Fig.B       

50µl of 3ml crude lentivirus with medium was directly used to infect 293T cells (without polybrene) grown in a separate 6 well plate. 48 hours later the expression of GFP was assessed.

Fig. C and Fig. D (magnification 100x): shRNA expression lentivirus was produced by the pLenti™ Lentivirus Production Kit (LT1001); and infected to HUVEC (Human Umbilical Vein Endothelial Cells).

Primary human umbilical vein endothelial cells (HUVEC) growing in 100mm dishes were infected by lentivirus shRNA negative control co-expressing with GFP. GFP expression were observed 3 days after infection.

Lentivirus-infection-in-human-endothelial-cells1-3br.jpg Fig.C

    Lentivirus-infection-in-human-endothelial-cells1-3fl.jpg Fig.D
   Fig.E (magnification 100x)            Lentivirus-in-vivo-example(fat-injection)                          

Fig.E: In vivo transduction sample

50 µl of purified lentivirus with our kit was injected into a mouse subcutaneous adipose tissue (without polybrene). After three days, the tissue was harvested and observed under fluorescence microscope and bright field.

                                        

 

Kit Components and Storeage Conditions 

 Catalog     Components         Size       Storage Condition    
LT1001-1 Lentivirus production medium   55ml
  4 °C. Add 1.2ml FBS before use.
LT1001-2 Helper-plasmids mixture   0.2ml
  4 °C
LT1001-3 Production helper-solution   0.5ml
  4 °C
LT1001-4 Plate coating solution   2.0ml
  4 °C
LT1001-5 Lentivirus Purification Solution   20ml
  4 °C
LT1001-6 Lentivirus storage solution   2.0ml
  4 °C
LT1001-7 GFP expressing lentivirus plasmid   1µg in 20µl TE   4 °C. Ampicillin resistance

 

References

1. Y Ido, et al. PLoS ONE, 2012 Apr 07( 4): e35092
2. Lan F, et. al. J Biol Chem. 2008 Oct 10;283(41):27628-35
 

Related Products

pLSLIC-PuroP2ATM (pLenti-SLIC-Puro-P2A, LT2002)    

pLSLIC-GfpP2ATM(pLenti-SLIC-GFP-P2A LT2003)

Pre-made shRNA Lentivirus Plasmids

Custom-made shRNA Lentivirus Plasmids

Pre-made shRNA expression lentivirus Particle

Custom-made shRNA expression lentivirus Particle

VP-Easy™ Chemical Competent Cell Preparation Kit

SLIC-Easy™ Chemical Competent Cell Preparation Kit

Omni-Easy™ Chemical Copmpetent Cell Kit

 

FAQs

Product Tags

Use spaces to separate tags. Use single quotes (') for phrases.