SRE TurboRFP Reporter Lentivirus (Hygromycin)

Catalog #

82310

$875 *

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

SRE TurboRFP Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain an TurboRFP (red fluorescent protein) reporter driven by the SRE (Serum Response Element) located upstream of the minimal TATA promoter. The lentiviruses also transduce a hygromycin selection gene (Figure 1). After transduction, activation of the MAPK (mitogen-activated protein kinase)/ERK (extracellular regulated kinase) signaling pathway in target cells can be monitored by assessing RFP expression.

gure 1. Schematic of the lenti-vector used to generate the SRE TurboRFP Reporter Lentivirus.

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Product Data Gallery

SRE TurboRFP reporter activity stimulated by human EGF or FBS in HEK293 cells transduced with SRE TurboRFP Reporter Lentivirus.

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of SRE TurboRFP Reporter Lentivirus at ≥10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
HEK293 cells	ATCC #CRL-1573
Thaw Medium 1	BPS Bioscience #60187
Assay Medium 1B	BPS Bioscience #79617
EGF Recombinant	BPS Bioscience #90201
FBS (Fetal Bovine Serum)	Invitrogen #26140-079
Lenti-Fuse™ Polybrene Viral Transduction Enhancer	BPS Bioscience #78939
96-well tissue culture, clear-bottom, white plate	Corning, #3610

Formulation

The lentiviruses were produced from HEK293T cells. Supplied in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations and produced at higher titers by special request, for an additional fee.

Background

The MAPK (mitogen-activated protein kinase)/ERK (extracellular regulated kinase) signaling pathway is a major participant in the regulation of cell growth and differentiation. It can be activated by various extracellular stimuli including mitogens, growth factors, and cytokines. Upon stimulation, MEK1/2 (MAP kinase kinase 1/2) phosphorylate and activate ERK1/2. The activated ERK1/2 translocate to the nucleus where they phosphorylate and activate transcription factors. The TCFs (Ternary Complex Factors), including transcription factor Elk1 (ETS like-1 protein), are among the best-characterized substrates of ERK. When phosphorylated by ERK, Elk1 forms a complex with Serum Response Factor (SRF) and binds to the Serum Response Element (SRE), resulting in the expression of numerous mitogen-inducible genes. TurboRFP, a red (orange) fluorescent protein derived from the sea anemone Entacmaea quadricolor, has high photostability and pH stability allowing for easy assay readouts.

References

Wong K.K., 2009 Recent Pat Anticancer Drug Discov. 4(1):28-35.
Treisman R., 1992 Trends Biochem Sci. 17(10): 423-426.

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.

Applications

Screen for activators or inhibitors of MAPK/ERK signaling pathway in transduced cells.
Generate SRE TurboRFP reporter cell pools or stable cell lines following hygromycin selection.

Shipping Temperature

-80°C

Notes

To generate an SRE-TurboRFP reporter cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of hygromycin (as pre-determined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Biosafety
The lentiviruses are produced with the SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. 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.

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only

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SRE TurboRFP Reporter Lentivirus (Hygromycin)

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