The RIK-2 monoclonal antibody specifically binds to CD253 which is also known as TRAIL (TNF-Related Apoptosis-Inducing Ligand) and Apo2 ligand (APO-2L). CD253 is a member of the Tumor Necrosis Factor Superfamily and is encoded by the TNFSF10 gene. CD253 is a type II membrane protein that may be expressed as a full-length, cell surface associated protein as well as in a soluble form. Both surface and soluble forms of TRAIL rapidly induce apoptosis by a wide range of cell lines but not by normal cells. TRAIL is expressed by activated T cells, NK cells, dendritic cells, monocytes and by a variety of non-lymphoid cells. TRAIL can bind to and exert apoptosis through DR4 (TRAIL-R1) and DR5 (TRAIL-R2) receptors. It can also bind to decoy receptors, including DcR1/TRID/TRAIL-R3 and DcR2/TRUNDD/TRAIL-R4, and possibly OPG/TNFRSF11B, which may serve to regulate TRAIL activity. TRAIL has been shown to be involved in T cell-mediated cytotoxicity, however its mechanism of action remains to be fully elucidated. The RIK-2 clone was selected based on its ability to block TRAIL-mediated cytotoxic activity. Flow Cytometric Analysis of TRAIL-induced Killing and RIK-2 Blocking using FITC Annexin V staining. Jurkat T cells were left untreated (far left/first panel) or treated for 16 hours under the following conditions: Cells were incubated with 20 ng/mL of recombinant human TRAIL and 10 μg/mL anti-histidine antibody (second panel), or with 20 ng/mL of recombinant human TRAIL and 10 μg/mL anti-histidine antibody preincubated with RIK-2 antibody (third panel), or with 20 ng/mL of recombinant human TRAIL and 10 μg/mL anti-histidine antibody preincubated with mIgG1 antibody (negative control) (far right/fourth panel). Following treatments, cells were evaluated for Annexin-V staining. Cells induced to undergo apoptosis by treatment with recombinant human TRAIL gave a population of cells that was Annexin V-FITC positive (second panel, M2). Annexin V-FITC staining was blocked when cells were incubated with a mixture of recombinant human TRAIL and RIK-2 antibody (third panel, M2). Cells treated with the mixture of recombinant TRAIL and mIgG1 antibody could not block the killing (far right/fourth panel). A small number of Annexin-V positive cells in the untreated population represents a basal level of apoptosis (far left/first panel). The results indicate that clone RIK-2 can block cell mediated killing induced by recombinant human TRAIL as measured by Annexin V-FITC staining of Jurkat cells.
Optimal working dilution should be determined by the investigator.
Beschränkungen
Nur für Forschungszwecke einsetzbar
Konzentration
1.0 mg/mL
Buffer
No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm sterile filtered.
Konservierungsmittel
Without preservative
Handhabung
This preparation contains no preservatives, thus it should be handled under aseptic conditions.
Lagerung
4 °C
Informationen zur Lagerung
Store undiluted at 4°C. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
Kayagaki, Yamaguchi, Nakayama, Kawasaki, Akiba, Okumura, Yagita: "Involvement of TNF-related apoptosis-inducing ligand in human CD4+ T cell-mediated cytotoxicity." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 162, Issue 5, pp. 2639-47, (1999) (PubMed).
Marsters, Pitti, Donahue, Ruppert, Bauer, Ashkenazi: "Activation of apoptosis by Apo-2 ligand is independent of FADD but blocked by CrmA." in: Current biology : CB, Vol. 6, Issue 6, pp. 750-2, (1997) (PubMed).
Sheridan, Marsters, Pitti, Gurney, Skubatch, Baldwin, Ramakrishnan, Gray, Baker, Wood, Goddard, Godowski, Ashkenazi: "Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors." in: Science (New York, N.Y.), Vol. 277, Issue 5327, pp. 818-21, (1997) (PubMed).
Pitti, Marsters, Ruppert, Donahue, Moore, Ashkenazi: "Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family." in: The Journal of biological chemistry, Vol. 271, Issue 22, pp. 12687-90, (1996) (PubMed).
Wiley, Schooley, Smolak, Din, Huang, Nicholl, Sutherland, Smith, Rauch, Smith: "Identification and characterization of a new member of the TNF family that induces apoptosis." in: Immunity, Vol. 3, Issue 6, pp. 673-82, (1996) (PubMed).
Target
TRAIL (TNFSF10)
(Tumor Necrosis Factor (Ligand) Superfamily, Member 10 (TNFSF10))