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RediFect Lentiviral Particles
Caution: For Laboratory Use. A product for research purposes only.

Name: RediFect Red-FLuc-Puro
Product Number: CLS960002

DESCRIPTIONCONTENTS Each vial contains 1 x 107 units/ml of lentiviral particles
resuspended in 200 ul of phosphate buffered saline.
 The packaged material provides sufficient number of lentiviralparticles for transduction of at least one cell line.

Noarmalized Intensity

RediFect Red-FLuc-Puro Lentiviral Particles are self-inactivating, recombination incompetent lentiviral particles carrying red-shifted LuciolaItalica luciferase transgene under control of the stable UbC promoter. The luciferase transgene is fused to the puromycin resistance gene via
T2A “self-cleaving” linker peptide for efficient co-expression with selection marker. The lentiviral particles are pseudotyped with G
glycoprotein from Vesicular Stomatitis Virus (VSVG), allowing efficient transduction of a wide variety of mammalian cells including mostcancer cell lines, primary, stem and non-dividing cells.

1008060
40200400

STORAGE & HANDLING Upon receipt, RediFect Red-FLuc-Puro Lentiviral Particles should
be IMMEDIATELY STORED AT -80 °C.
 When stored and handled properly, RediFect Red-FLuc-Puro
Lentiviral Particles are stable for three months.
 After thawing the vial, gently spin down the solution and place it onice. Avoid repeated freeze-thawing as this will reduce viral titer.

500
600
700
Wavelength (nm)800

Normalized bioluminescence emission spectra of MDA-MB231 cell linetransduced with red-shifted luciferase from Luciola Italica

APPLICATIONS In vitro labeling of mammalian tumor cells for non-invasive in vivo monitoring of tumor growth in subcutaneous and orthotopic deep tissue
mouse models
 In vitro labeling of stem cells for non-invasive in vivo monitoring of survival and growth of implanted cells.

NOTES
 For laboratory use only. This product is intended for animal research only and not for use in humans. It must be used by or directly under
the supervision of a technically qualified individual experienced in handling potentially hazardous materials. Please read the Material
Safety Data Sheet (MSDS) provided for this product.
 Several of PerkinElmer’s products and product applications are covered by U.S and foreign patents and patents pending. Our products
are not available for resale or other commercial uses without a specific agreement from PerkinElmer.

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Protocol for the use of RediFect Red-FLuc-Puromycin Lentiviral Particles
1) Day 1. Plate 20,000-50,000 of mammalian cells in complete medium into 1 well of a 24 well-plate. Incubate cells for 12-24hr.
2) Day 2. Thaw a vial with lentiviral particles, gently spin down the solution and place it on ice. Replace the medium with 500ul of freshcomplete medium containing hexadimethrine bromide (polybrene) at a final concentration of 4 ug/ml. Add the appropriate amount of viralparticles at a suitable multiplicity of infection (MOI) directly to cells. Incubate cells with the viral particles for 24 hr.
a. Polybrene enhances transduction of most cell lines. However, some cells such as mesenchymal stem cells or primaryneurons are known to be sensitive to polybrene. If the cells are sensitive, do not add polybrene and the cells should stillbe transduced.
b. If the optimal MOI is unknown for the cell line of choice, it is recommended to use a range of MOIs (20-100).
3) Day 3. Replace the virus containing medium with 1ml of fresh pre-warmed complete culture medium and incubate cells for 24hr.
4) Day 4. Spit the cells if necessary; add fresh complete culture medium with the appropriate amount of puromycin to select the transduced
cells. A kill curve experiment is recommended when the appropriate amount of puromycin is unknown for the cell line.
5) Day 6 and forward. Expand puromycin resistant cells to assay for expression of luciferase.

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