AP1 eGFP Reporter Lentivirus (JNK Signaling Pathway)

Catalog #

78680

$875 *

Size: 500 µl x 2

Qty

Bulk/Custom

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Description

The Activator protein 1 (AP1) eGFP Reporter Lentiviruses are replication incompetent, HIV-based, 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 eGFP gene driven by an AP1 response element located upstream of the minimal TATA promoter (Figure 1). After transduction, activation of the JNK signaling pathway and AP1-mediated activity in the target cells can be monitored by eGFP expression.

Figure 1. Schematic of the lenti-vector used to generate the AP1-eGFP Reporter Lentivirus.

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Synonyms

JNK-1, JNK-2

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Product Data Gallery

AP1 eGFP reporter expression stimulated by PMA in HEK293 cells

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 will be provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
Phorbol 12-Myristate 13-Acetate (PMA)	LC Laboratories #P-1680
HEK293 growth medium or Thaw Medium 9	BPS Bioscience #79665
Assay Medium 1B	BPS Bioscience #79617
Polybrene	Millipore #TR-1003-G

Formulation

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

Background

The stress-activated protein kinase/c-jun N-terminal kinase (SAPK/JNK) family of proteins are activated by stress, inflammatory cytokines, mitogens, oncogenes, and inducers of cell differentiation and morphogenesis. Upon activation of the SAPK/JNK pathway, MAP Kinase Kinases phosphorylate and activate JNKs. The activated JNKs translocate to the nucleus where they phosphorylate and activate transcription factors such as c-Jun, c-Fos, ATF (Activating transcription factor), and JDP (Jun dimerization protein). These transcription factors form heterodimers termed AP1, which bind to AP1 response elements in the promoters of target genes and induce their transcription.

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 the JNK signaling pathway in the transduced target cells.
Generate an AP1 eGFP Reporter stable cell line (geneticin-resistant).

Shipping Temperature

-80°C

Notes

To generate the AP1 eGFP reporter stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of G418 for antibiotic selection of transduced cells. For information about determining the appropriate concentration of selection antibiotic, visit https://bpsbioscience.com/cell-line-faq in our Resources section (Q: what is a kill curve?)

Biosafety
The lentiviruses are produced with the third generation 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 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

Cell Signaling Pathway

Regulatory Status

Research Use Only

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AP1 eGFP Reporter Lentivirus (JNK Signaling Pathway)

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