GAL4 DBD-GR Lentivirus

Catalog #

78632

$795 *

Size: 500 µl x 2

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Bulk/Custom

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

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Description

GAL4 DBD-GR (GAL4 DNA binding domain-glucocorticoid receptor) 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 express a fusion protein in which the glucocorticoid receptor (GR) ligand binding domain is fused to the DNA binding domain of GAL4 (GAL4 DBD) (Figure 1). After co-transduction of GAL4 DBD-GR lentivirus and GAL4-responsive UAS luciferase reporter lentivirus (BPS Bioscience #78631), the glucocorticoid-induced activation of the glucocorticoid receptor can be monitored by measuring the luciferase activity.

Figure 1. Schematic of the lenti-vector used to generate the GAL4 DBD-GR Lentivirus.

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Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer ≥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	ATCC #CRL-1573
Thaw Medium 1	BPS Bioscience #60187
UAS luciferase reporter lentivirus	BPS Bioscience #78631
Dexamethasone	Sigma #D4902
Polybrene Infection / Transfection Reagent	Millipore #TR-1003-G
96-well Clear Bottom White Polystyrene TC-treated	Corning #3610
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690
Luminometer

Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.

Background

The glucocorticoid signaling pathway plays an important role in development, fluid homeostasis, cognition, immune response and metabolism. Glucocorticoids are a class of steroid hormones that bind to the glucocorticoid receptor, causing it to translocate to the nucleus. Upon translocation, the receptor can regulate the transcription of many genes, including those that regulate glucose metabolism and inflammatory responses.

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

In combination with UAS Luciferase reporter lentivirus (BPS Bioscience #78631), to monitor glucocorticoid signaling pathway activity.
Generate a GAL4 DBD-GR stable cell line (Hygromycin resistant).

Shipping Temperature

-80°C

Notes

To generate a GAL4 DBD-GR stable 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) for antibiotics selection of transduced cells.

Biosafety
The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after 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 and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

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

Warnings

Avoid freeze/thaw cycles

Scientific Category

Primary/Replication Incompetent

Regulatory Status

Research Use Only

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GAL4 DBD-GR Lentivirus

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