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5452
Human His6Fas Ligand/TNFSF6 (hHis6FasL)
Cytokines & Growth Factors
Growth Factors and Cytokines

Human His6Fas Ligand/TNFSF6 (hHis6FasL) #5452

Citations (0)
The viability of Jurkat cells treated with increasing amounts of hHis6FasL in the presence of 10 µg/ml anti-His antibody was assessed. After 24 hour treatment with hHis6FasL, cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.
The purity of recombinant hHis6FasL was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hHis6FasL and staining overnight with Coomassie Blue.
Western blot analysis of extracts from Jurkat cells, untreated or treated with hHis6FasL for 3 hours, using Cleaved PARP (Asp214) Antibody (Human Specific) #9541 (upper) or PARP Antibody #9542 (lower).
Treatment of Jurkat cells with hHis6FasL induces casapase-3 cleavage as detected by PathScan® Cleaved Caspase-3 (Asp175) Sandwich ELISA Kit #7190.

Product Usage Information

Reconstitution:

With carrier: Add sterile PBS or PBS containing 1% bovine or human serum albumin or 5-10% FBS to a final hHis6FasL concentration of greater than 50 μg/ml. Solubilize for 30 minutes at room temperature with occasional gentle vortexing.

Carrier free: Add sterile PBS or PBS containing protein to minimize absorption of hHis6FasL to surfaces. Solubilize for 30 minutes at room temperature with occasional gentle vortexing. Stock hHis6FasL should be greater than 50 μg/ml.

Storage

Stable in lyophilized state at -20°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.
Maintain sterility. Storage at -20°C should be in a manual defrost freezer.

Product Description

MW (kDa) 31-36
Purity >98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hHis6FasL. All lots are greater than 98% pure.
Endotoxin Less than 0.01 ng endotoxin/1 μg hHis6FasL.
Activity The bioactivity of hHis6FasL was determined in a Jurkat cell viability assay. The ED50 of each lot is between 1-5 ng/ml.
Molecular Formula Recombinant N-terminally His6-tagged hFasL has a calculated MW of 19,834. DTT-reduced and non-reduced protein migrate as 31-36 kDa polypeptides. Lower mobility in SDS-PAGE is due to glycosylation.

Source / Purification

Recombinant humanHis6 FasL (hHis6FasL) Pro134-Leu281 (Accession #NP_000630) was expressed in human 293 cells at Cell Signaling Technology.

Background

FasL is a member of the TNF-superfamily family of proteins and is expressed primarily on the cell surface of activated T and NK cells (1). FasL regulates the immune response through its ability to induce apoptosis. The immunoregulatory role of FasL is underscored by lymphoadenopathy associated with FasL or Fas knockout mice and the fraction of autoimmune lympho-proliferative syndrome (ALPS) patients that have mutations in the FasL receptor, Fas (1). FasL is a membrane protein that can be cleaved into a soluble trimeric form by metalloproteinases (1,2). The soluble form of FasL retains the ability to bind to Fas, however, its ability to induce apoptosis is diminished (2). The ligation of Fas by FasL leads to the assembly of death-inducing signaling complex (DISC) and the recruitment and activation of caspase-8/caspase-10 (1,3). Active caspase-8/caspase-10 subsequently activates the “effector” caspases caspase-3 and caspase-7, and cleavage of BID (1,3).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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