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Cell Immortalization Reagents

Primary cells undergo a finite number of population doublings and reach a state called replicative senescence after limited cell divisions. GeneCopoeia’s Cell Immortalization Reagents use recombinant lentivirus carrying genes, such as the simian virus 40 (SV40) T antigen, the human telomerase reverse transcriptase protein (hTERT), CDK4, HOXB, HOXA, cMyc, Bmi1 and HPV-16 E6/E7 to induce immortalization and significantly prolong primary cells' lifespan.

 

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Description                                              

Price
LP721-025 CMV  Puromycin

SV40 small T & large T antigen

Purified Lentiviral Particles (25 µl x 1 vial)

$295

LP721-100    

  CMV      Puromycin

SV40 small T & large T antigen     

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP722-025 EF1α Puromycin

SV40 small T & large T antigen

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP722-100 EF1α Puromycin

SV40 small T & large T antigen      

Purified Lentiviral Particles (25 µl x 4 vial)

$725

LP742-025 CMV Puromycin

SV40 large T antigen                         

Purified Lentiviral Particles (25 µl x 1 vial)

$295

LP742-100

CMV Puromycin

SV40 large T antigen                         

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP724-025 CMV N/A

SV40 large T antigen                         

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP724-100 CMV N/A

SV40 large T antigen                         

Purified  Lentiviral Particles (25 µl x 4 vial)

$725
LP725-025 EF1α N/A

SV40 large T antigen                         

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP725-100 EF1α N/A

SV40 large T antigen                         

Purified  Lentiviral Particles (25 µl x 4 vial)

$725
LP743-025 EF1α Puromycin

SV40 large T antigen                         

Purified  Lentiviral Particles (25 µl x 1 vial)

$295
LP743-100 EF1α Puromycin

SV40 large T antigen                          

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP723-025 EF1α Hygromycin

SV40 large T antigen                     

Purified  Lentiviral Particles (25 µl x 1 vial)

$295
LP723-100 EF1α Hygromycin

SV40 large T antigen                         

Purified  Lentiviral Particles (25 µl x 4 vial)

$725
LP726-025 CMV Puromycin

hTERT                                                 

Purified  Lentiviral Particles (25 µl x 1 vial)  

$295
LP726-100 CMV Puromycin

hTERT                                                  

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP727-025 EF1α Hygromycin

hTERT

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP727-100 EF1α Hygromycin

hTERT

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP728-025 CMV Hygromycin

hTERT                                                  

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP728-100 CMV Hygromycin

hTERT

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP729-025 CMV N/A

hTERT

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP729-100 CMV N/A

hTERT

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP730-025 EF1α Puromycin

hTERT

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP730-100 EF1α Puromycin

hTERT

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP744-025 EF1α N/A

hTERT

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP744-100 EF1α N/A

hTERT

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP731-025 CMV Puromycin

CDK4

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP731-100 CMV Puromycin

CDK4

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP732-025 CMV N/A

CDK4

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP732-100 CMV N/A

CDK4

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP733-025 CMV Puromycin

HOXB8

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP733-100 CMV Puromycin

HOXB8

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP734-025 CMV N/A

HOXB8

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP734-100 CMV N/A

HOXB8

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP735-025 CMV Puromycin

HOXA9

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP735-100 CMV Puromycin

HOXA9

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP736-025 CMV N/A

HOXA9

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP736-100 CMV N/A

HOXA9

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP738-025 CMV N/A

HOXA10

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP738-100 CMV N/A

HOXA10

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP737-025 CMV Puromycin

HOXA10

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP737-100 CMV Puromycin

HOXA10

Purified Lentiviral Particles (25 µl x 4 vial)

$725

LP741-025 CMV Puromycin

c-Myc

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP741-100 CMV Puromycin

c-Myc

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP739-025 CMV Puromycin

Bmi1

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP739-100 CMV Puromycin

Bmi1

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP740-025 CMV N/A

Bmi1

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP740-100 CMV N/A

Bmi1

Purified Lentiviral Particles (25 µl x 4 vial)

$725
LP745-025 CMV Puromycin

HPV-16 E6-E7

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP745-100 CMV Puromycin

HPV-16 E6-E7

Purified Lentiviral Particles (25 µl x 4 vials)

$725
LP746-025 CMV N/A

HPV-16 E6-E7

Purified Lentiviral Particles (25 µl x 1 vial)

$295
LP746-100 CMV N/A

HPV-16 E6-E7

Purified Lentiviral Particles (25 µl x 4 vials)

$725

 

 

 

GeneCopoeia Lentifect™ Lentiviral Particles are produced from a standardized protocol using purified plasmid DNA and the proprietary reagents EndoFectin™ Lenti (for transfection) and TiterBoost™ solution. The protocol uses a third-generation self-inactivating packaging system meeting BioSafety Level 2 requirements.

The Lentifect™ particles include CMV or EF1α promoter for efficient expression of the immortalization inducer gene in target cells with puromycin or hygromycin resistance marker for selection of stably transduced cells. The immortalization lentivirus particles without antibiotic selection marker are also available.

Provided as 4 vials of 25 µl of lentiviral particles with titers of ~1 x 108 TU/ml. Lentifect™ particles are shipped on dry ice and must be stored at –80°C immediately upon receipt. Avoid repeated freeze-thaw cycles as this will reduce titers.

The lentiviral expression construct was validated by full-length sequencing, restriction enzyme digestion and PCR-size validation using gene-specific and vector-specific primers. The product is confirmed free of bacteria, fungi and common Mycoplasma contamination.

 

Cell Immortalization Protocol

  • Plate target primary cells in a 6-well plate at density of 0.5-2 x105 cells per well
  • Thaw the lentiviral particles in 37 ºC water bath and immediately remove it from the water bath once thawed
  • Transduce the cells in a 6-well plate with immortalizing lentivirus or GFP-lentivirus (positive control) at a MOI of 10-100. Dilute the lentivirus with the fresh complete medium if cytotoxicity is a problem
  • The next day, remove viral supernatant and add appropriate complete growth medium to the cells and incubate at 37 ºC
  • After 72 hours incubation, passage the transduced cells in complete medium supplemented with the appropriate amount of selection antibiotic
  • Pick antibiotic-resistant colonies and cultivate continuously in complete medium with antibiotics to test the immortalization