Notch1dE Lentivirus

Catalog #

78747

$885 *

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

Notch1dE Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most mammalian cells, including primary and non-dividing cells. These lentiviruses express a truncated human Notch1 construct (Notch1dE) in which the entire extracellular domain was deleted. The lentiviruses also contain a puromycin selection marker (Figure 1). Once expressed in a cell line of interest, Notch1dE is cleaved by γ-secretase, resulting in the constitutively active intracellular domain of Notch (NICD). NICD translocates to the nucleus, binds to the transcription factor CSL (CBF1/RBPJκ/Suppressor of Hairless/Lag-1) and activates transcription of Notch1-responsive genes.

When used in combination with CSL (CBF1/RBP-Jk) Luciferase Reporter Lentiviruses (BPS Bioscience #78746), NICD activates the CSL (CBF1/RBPJκ/Suppressor of Hairless/Lag-1) responsive elements, inducing luciferase expression.

Figure 1. Schematic of the lenti-vector used to generate the Notch1dE lentivirus.

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Synonyms

Notch-1

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Product Data Gallery

CSL driven luciferase reporter activity in HEK293 cells transduced with CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus and/or Notch1dE Lentivirus.

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer ≥10⁷ TU/ml. The titer varies 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
CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus	BPS Bioscience #78746
96-well tissue culture, clear-bottom, white plate	Corning #3610
One-Step™ Luciferase Assay System	BPS Bioscience #60690
Luminometer

Formulation

The lentivirus particles were produced in HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.

Background

The Notch signaling pathway controls cell fate decisions in vertebrates and invertebrates’ tissues and is involved in embryonic development, tissue homeostasis, and regulation of the immune and angiogenic systems. Notch signaling is triggered through the binding of a transmembrane ligand, present in opposing cells, to one of the four existing Notch transmembrane receptors (Notch1/Notch2/Notch3/Notch4). This results in proteolytic cleavage of the Notch receptor, releasing the constitutively active intracellular domain of the Notch receptor (NICD). NICD translocate to the nucleus and associates with the transcription factor CSL (CBF1/RBPJκ/Suppressor of Hairless/Lag-1) and coactivator Mastermind to turn on the transcription of Notch-responsive genes. Dysfunction of Notch signaling has severe consequences, including developmental pathologies or cancer (such as T cell acute lymphoblastic leukemia, T-ALL, and urothelial bladder cancer). The use of Notch inhibitors, mainly gamma-secretase inhibitors, as a cancer therapy option and in the regeneration of tissues is ongoing. Further studies will allow us to have a deeper understanding of Notch signaling and will benefit future therapeutic approaches.

References

Lu F.M., et al., 1996 Natl. Acad. Sci. USA 93(11): 5663-5667.
Kanungo J., et al., 2008 Neurochem. 106: 2236-48.
Cao L. et al., 2023 Blood Adv. 1182/bloodadvances.2023010380

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.

Growth Media

For best results, the use of validated and optimized media from BPS Bioscience is highly recommended. Other preparations or formulations of media may result in suboptimal performance.

Media Required for the Proposed Assay

Thaw Medium 1 (BPS Bioscience #60187): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin.

Applications

Monitor Notch signaling pathway activity, when used in combination with the CSL (CBF1/RBP-Jk) Luciferase Reporter Lentivirus (BPS Bioscience #78746).
Generate Notch1dE-expressing cell pools or stable cell lines by puromycin selection.

Shipping Temperature

-80°C

Notes

To generate a Notch1dE stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve, FAQs (bpsbioscience.com)) for antibiotic selection of transduced cells, followed by clonal selection.

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

Cell Signaling Pathway

Regulatory Status

Research Use Only

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Notch1dE Lentivirus

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