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RANKL (soluble) (human), (recombinant)

 
ALX-522-012-C010 10 µg 396.00 USD
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Product Specification

Alternative Name:TRANCE, OPGL, ODF, TNFSF 11, CD254
 
MW:~18kDa (non-glycosylated), ~28kDa (glycosylated) (SDS-PAGE).
 
Source:Produced in HEK 293 cells. The extracellular domain of human RANKL (aa 152-317) is fused at the N-terminus to a linker peptide (6 aa) and a FLAG®-tag.
 
UniProt ID:O14788
 
Concentration:0.1mg/ml after reconstitution.
 
Formulation:Lyophilized. Contains PBS.
 
Purity:≥90% (SDS-PAGE)
 
Endotoxin Content:<0.1EU/µg purified protein (LAL test; Associates of Cape Cod).
 
Species reactivity:Human, Mouse
 
Specificity:Binds to human and mouse RANK.
 
Applications:ELISA
 
Application Notes:ELISA: binds to RANK receptor.
 
Solubility:Soluble in water.
 
Reconstitution:Reconstitute with 100µl sterile water. Further dilutions should be made with medium containing 5% fetal calf serum or a carrier protein.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
Use/Stability:Stable for at least 6 months after receipt when stored at -20°C.
 
Handling:Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -20°C.
 
Scientific Background:RANK ligand (RANKL), also called TRANCE, OPGL and ODF, is a member of the TNF family that is expressed in activated T cells, lymph nodes and in stromal cell lines. RANKL (TRANCE) interacts with its receptor RANK expressed on mature dendritic cells (DC) and mature osteoclasts, leading to the inhibition of apoptosis, probably through the upregulation of bcl-x. RANKL is, therefore, a useful tool for enhancing DC and osteoclast survival and activity. Osteoprotegerin (OPG, see Prod. No. ALX-522-007) is a potent inhibitor of RANKL.
Historical data has shown that the biological activity of RANKL supports the survival of dendritic cells and osteoclasts.
 
Technical Info/Product Notes:FLAG is a registered trademark of Sigma-Aldrich Co.
 
522.012-1
Figure 1: Schematic structure of human rhsRANKL (peptide, aa. 151-316).
522.012-2
Figure 2: Binding of rhsRANKL to Osteoprotegerin. Recombinant, human sRANKL binds to Osteoprotegerin:Fc (see Prod. No. ALX-522-007).Method: Ligand binding assay: 96 well ELISA plates were coated O/N with 50 ng OPG-Fc (Prod. No. ALX-522-007) per well. After a blocking step, the indicated concentrations of Flag-tagged RANKL were added for 1 hour. Bound ligand was revealed with an anti-FLAG antibody (Prod. No. ALX-804-034) at 1 µ/ml for 30 minutes, followed by a rabbit anti-mouse IgG-HRP at 1/1000 dilution for 30 minutes. OPD was used as a substrate for the peroxidase. Absorbance was measured at 490 nm in an ELISA reader.
522-012 3
Figure 3: Mononuclear cells differentiate into Osteoclasts in the presence of M-CSF and RANKL, Soluble (human) (recombinant) (ALX-522-012). Method: Human CD14+ mononuclear cells isolated from adult peripheral blood were cultured in control medium (upper panel), in 25ng/ml M-CSF (middle panel), or in 25ng/ml M-CSF plus 50ng/ml RANKL (lower panel). Osteoclasts were identified as Tartrate-Resistant Acid Phosphatase (TRAP)-positive multinucleated cells. Osteoclasts were detected exclusively in presence of RANKL and in these culture conditions (M-CSF + RANKL), cells fused and generated multinucleated dark red TRAP-positive cells (arrows). Nuclei were stained with haematoxylin. Original magnification: 200x. Picture courtesy of V.Kindler and D.Suvà, University of Geneva.
522-012 SDS-PAGE
SDS-PAGE Analysis of RANKL (soluble) (human), (recombinant). Lane 1: MW Marker, Lane 2: 1µg.
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522.012-1 522.012-2 522-012 3 522-012 SDS-PAGE

Product Literature References

A novel in vivo role for osteoprotegerin ligand in activation of monocyte effector function and inflammatory response: D. Seshasayee, et al.; J. Biol. Chem. 279, 30202 (2004), Abstract;
Induction of Cell Cycle Arrest and Apoptosis by the Proteasome Inhibitor PS-341 in Hodgkin Disease Cell Lines Is Independent of Inhibitor of Nuclear Factor-kappaB Mutations or Activation of the CD30, CD40, and RANK Receptors: B. Zheng, et al.; Clin. Cancer Res. 10, 3207 (2004), Abstract;
Suppression of IL-12 Production by Soluble CD40 Ligand: Evidence for Involvement of the p44/42 Mitogen-Activated Protein Kinase Pathway: M. Wittmann, et al.; J. Immunol. 168, 3793 (2002), Abstract;
Functional expression of receptor activator of nuclear factor kappaB in Hodgkin disease cell lines: P. Fiumara, et al.; Blood 98, 2784 (2001), Abstract; Full Text
Receptor activator of NF-κB and osteoprotegerin expression by human microvascular endothelial cells, regulation by inflammatory cytokines, and role in human osteoclastogenesis: P. Collin-Osdoby, et al.; J. Biol. Chem. 276, 20659 (2001), Abstract; Full Text

General Literature References

Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL: H. Yasuda, et al.; PNAS 95, 3597 (1998), Abstract;
Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation: D.L. Lacey, et al.; Cell 93, 165 (1998), Abstract;
A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function: D.M. Anderson, et al.; Nature 390, 175 (1997), Abstract;
TRANCE (tumor necrosis factor [TNF]-related activation-induced cytokine), a new TNF family member predominantly expressed in T cells, is a dendritic cell-specific survival factor: B.R. Wong, et al.; J. Exp. Med. 186, 2075 (1997), Abstract;

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