The activity of hHis64-1BBL was assessed by its ability to bind to 4-1BB in a functional ELISA.
The purity of recombinant hHis64-1BBL was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hHis64-1BBL and staining overnight with Coomassie Blue.
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hHis64-1BBL. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Stable in lyophilized state at 4°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.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hHis64-1BBL. All lots are greater than 98% pure.
Recombinant N-terminally His6-tagged h4-1BBL has a calculated MW of 22,020. DTT-reduced and non-reduced protein migrate as 22 kDa polypeptides. The expected amino terminus of recombinant hHis64-1BBL was verified by amino acid sequencing.
The activity of hHis64-1BBL was assessed by its ability to bind to 4-1BB in a functional ELISA. The concentration at which half-maximal binding was observed for each lot was 6-18 ng/ml.
Less than 0.01 ng endotoxin/1 μg hHis64-1BBL.
Recombinant human His64-1BBL (hHis64-1BBL) Arg71-Glu254 (Accession #NP_003802) was expressed in human 293 cells at Cell Signaling Technology.
4-1BBL (also known as TNFSF9) is a member of the TNF-R superfamily, which includes OX40L, CD27L, CD40L, and GITRL (1). 4-1BBL is expressed primarily on antigen presenting cells, such as macrophages, dendritic cells, and B cells (2). 4-1BBL binds to 4-1BB on the surface of activated T cells (1). 4-1BBL/4-1BB binding activates the NF-κB pathway via TRAF1, 2, and 3 (1). 4-1BBL functions as a co-stimulatory molecule, inducing T cell proliferation, cytolytic activity, and cytokine production (2). Matrix metalloproteases can generate soluble 4-1BBL by cleaving it from the cell surface (3). The in vitro activity of soluble 4-1BBL is enhanced by cross-linking (4,5).
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