YFP (Topaz) Lentivirus

Catalog #

79989

$795 *

Size: 500 µl x 2

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

Topaz1, a yellow fluorescence protein (YFP), is one of the brightest and thermally stable green fluorescence proteins (GFP). The YFP (Topaz) Reporter Lentivirus contains replication-defective, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain an EF1α promoter-driven YFP(Topaz) construct (a.a. sequence below), and confer both YFP (Topaz) expression and puromycin resistance to the target cells (Figure 1). YFP (Topaz) expression and transduction efficiency can easily be verified and optimized via fluorescence microscopy or flow cytometry. Topaz has an excitation wavelength of 514 nm, an emission wavelength of 527 nm, and extinction coefficient of 94,500M-1cm-1.

Figure 1. Schematic of the lentiviral vector used to generate the YFP (Topaz) reporter

Figure 1. Schematic of the lentiviral vector used to generate the YFP (Topaz) reporter

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Synonyms

79703, Topaz1, yellow fluorescence protein lentivirus, GFP lentivirus

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Product Data Gallery

Product Info

Storage and Usage

Citations

Supplied As

Two vials (2 x 500 μl) of YFP (Topaz) reporter lentivirus at a titer ≥5 x 10^6 TU/ml. The titer will vary with each lot, and multiplicity of infection will depend on the target cell type. We provide a standardized p24 measurement with each shipment

Materials Required But Not Supplied

• Thaw Medium 10 (BPS Bioscience #79704):RPMI 1640 medium (Thermo Fisher, #A1049101) supplemented with 10% FBS (Thermo Fisher, #26140079), 1 mM Sodium pyruvate (Corning, #25-000-CL), 1% Non-essential amino acids (Corning, #25-025-CL), 1% Penicillin/streptomycin (Thermo Fisher, #15140122)
• Recombinant Human IL-2 (BPS Bioscience, #90184)
• Polybrene (Millipore, #TR-1003-G)
• Puromycin (Invivogen, #ANT-PR-1)
• T cell activation reagents (αCD3/αCD28 or Phytohaemagglutinin (PHA))
• Recombinant Human IL-17 (BPS Bioscience, #91014) (optional)
• Recombinant Human IL-15 (BPS Bioscience, #90180) (optional)
• 6-well and T75 tissue culture-treated assay plates
• Flow cytometer or fluorescence microscope

Amino Acid Sequence

Variation of mVENUS, with L69V and K80R mutations

Genbank #

AUG68261

References

Cubitt, A.B. et. al., Understanding structure-function relationship in the Aequorea victoria green fluorescence protein, Methods in Cell Biology, 1998;58:19-30.

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.

Instructions for Use

See assay protocol for detailed instructions.

Shipping Temperature

-80°C

Notes

*Tip: Transduction efficiency can vary between lentivirus lots and different cell donors. While using a higher MOI/titer can provide limited improvement, we recommend optimizing this step by minimizing the overall infection volume and maximizing T cell density.

Warnings

Biosafety
None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only

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YFP (Topaz) Lentivirus

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