Membrane-Bound TNFα (mTNFα) Lentivirus

Catalog #

78955

$995 *

Size: 500 µl x 2

Qty

Bulk/Custom

Add to Favorites

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

●

Purchase

●

Description

Membrane-Bound TNFα Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These particles result in expression of uncleavable, membrane-bound TNFα (Tumor necrosis factor alpha) driven by an EF1a promoter, and a puromycin selection marker (Figure 1).

Figure 1. Schematic of the lenti-vector used to generate Membrane-Bound TNFα (mTNFα) Lentivirus.

●

Product Data Gallery

Expression of membrane-bound TNFα in CHO cells transduced with Membrane-Bound TNFα (mTNFα) Lentivirus by flow cytometry.

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
Thaw Medium 3	BPS Bioscience #60186
Lenti-Fuse™ Polybrene Viral Transduction Enhancer	BPS Bioscience #78939
12-well tissue culture-treated plates
Flow cytometer or fluorescence microscope

Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture 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

Tumor necrosis factor (TNF, also known as TNFα) is a cytokine produced predominantly by activated macrophages and T lymphocytes. It has been identified as a key regulator in inflammatory and immune responses. TNF signaling pathways are triggered by binding to one of two distinct receptors, designated TNFR1 (TNF receptor 1) and TNFR2, which are differentially regulated on various cell types in normal and diseased tissues. TNFα exists in both a trimeric membrane-bound form (mTNFα) and as a soluble protein. TNFα is synthetized in a precursor form, a cell surface type II transmembrane protein, which is cleaved by metalloproteinases such as TACE (TNFα converting enzyme) into a soluble peptide. Soluble TNFα can then bind to its receptors and activate downstream signaling pathways. Transmembrane TNFα can also bind to TNFα receptors and induce cellular responses. For instance, it can enhance cytotoxicity in NK cells, while in the liver it can trigger hepatitis. Anti-TNFα antibodies can bind to mTNFα and trigger antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) to destroy the mTNFα-expressing inflammatory cells, being a promising therapy for inflammatory diseases.

References

Wang F., et al., 2017 Mol Med Rep 16: 1021-1030.
Horiuchi T., et al., 2010 Rheumatology 49(7):1215-1228.

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 Cellular Assay
Thaw Medium 3 (BPS Bioscience #60186): F-12K medium supplemented with 10% FBS, 1% Penicillin/Streptomycin.

Applications

Generate cell pools or stable cell line expressing membrane-bound TNFα following puromycin selection.
Study anti-TNFα antibodies ADCC (antibody dependent cellular cytotoxicity) potential.

Shipping Temperature

-80°C

Notes

To generate a membrane-bound TNFα expressing 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, 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 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.

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cancer Therapy Target

Regulatory Status

Research Use Only

Skip to the end of the images gallery

Skip to the beginning of the images gallery

Membrane-Bound TNFα (mTNFα) Lentivirus

PRODUCTS

SERVICES

RESOURCES

COMPANY