Recombinant human RANKL protein (Active) (ab157289)
Key features and details
- Expression system: HEK 293 cells
- Purity: > 90% SDS-PAGE
- Endotoxin level:
- Active: Yes
- Tags: DDDDK tag N-Terminus
- Suitable for: ELISA, Functional Studies, SDS-PAGE
Preparation and Storage
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Stability and Storage
Shipped at 4°C. Store at -20°C. Avoid freeze / thaw cycle.
Constituent: PBS
This product is an active protein and may elicit a biological response in vivo, handle with caution.
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ReconstitutionReconstitute with 100µl sterile water to a concentration of 0.1 mg/ml. Further dilutions should be made with medium containing 5% fetal calf serum or a carrier protein. After reconstitution, prepare aliquots and store at -20°.
Images
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Functional Studies - Recombinant human RANKL protein (ab157289)Binding of ab157289 to Osteoprotegerin. ab157289 binds to Osteoprotegerin:Fc.
Method: Ligand binding assay: 96 well ELISA plates were coated O/N with 50 ng OPG-Fc per well. After a blocking step, the indicated concentrations of ab157289 were added for 1 hour. Bound ligand was revealed with an anti-FLAG antibody at 1 µg/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.
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SDS-PAGE - Recombinant human RANKL protein (ab157289)SDS-PAGE analysis of ab157289. Lane 1: MWt Marker, Lane 2: ab157289 1µg. -
Functional Studies - Recombinant human RANKL protein (ab157289)Mononuclear cells differentiate into Osteoclasts in the presence of M-CSF and ab157289.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 ab157289 (lower panel). Osteoclasts were identified as Tartrate-Resistant Acid Phosphatase (TRAP)-positive multinucleated cells. Osteoclasts were detected exclusively in presence of ab157289 and in these culture conditions (M-CSF + ab157289), cells fused and generated multinucleated dark red TRAP-positive cells (arrows). Nuclei were stained with haematoxylin.
