4-1BB Ligand is a type II transmembrane glycoprotein belonging to the TNF superfamily (TNFSF9). It is expressed on activated antigen presenting cells (APCs) and on most T and B lymphoma cell lines. Soluble 4-1BB Ligand can be released from the cell surface following cellular activation via proteolytic cleavage. 4-1BB Ligand exists as a noncovalent homotrimer and interacts with 4-1BB (TNFRSF9) to provide T cell costimulatory signals and dendritic cell activation.
Applications:
Suitable for use in Western Blot, Flow Cytometry and ELISA. Other applications not tested.
Recommended Dilution:
Western Blot: 1-2ug/ml. The detection limit is ~1ng/lane under non-reducing and reducing conditions.
Flow Cytometry: 25ug/ml. 10ul of the diluted solution labels 1-2.5 x 10e5 cells in a total reaction volume (same/less than) 200ul. The binding of unlabeled monoclonal antibodies may be visualized by adding 10ul of a 25ug/ml stock solution of a secondary developing reagent such as Gt xRt IgG conjugated to a fluorochrome.
Direct ELISA: 0.5-1.0ug/ml. The detection limit is ~1.5ng/well.
Optimal dilutions to be determined by the researcher.
Hybridoma:
Fusion of a mouse myeloma with B cells obtained from a rat immunized with purified, Sf 21-derived, recombinant mouse 4-1BB Ligand extracellular domain.
Storage and Stability:
Lyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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