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High sensitivity chemiluminescent readout (signal-to-noise ratio of ≥ 10)
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Rapid protocol yields results in just 2.5 hours
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No cell lines or transfection reagents required
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Reproducible, high-throughput amenable assay (Z’-factor 0.871)
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Amenable for high-throughput screening applications
The LEADING LIGHT® Sclerostin Screening System is a 1x96 well plate assay based on Sclerostin–LRP interaction. This system contains engineered LRP5-alkaline phosphatase (ALP) and human Sclerostin immobilized in a multi-well plate. LRP5-alkaline phosphatase binds to Sclerostin coated on the bottom of the well and is detected by the activity of alkaline phosphatase remaining after a washing step. This system can be used for drug screening to identify small molecule compounds, antibodies, DNA aptamers and peptides capable of modulating sclerostin-LRP interaction.
Inhibition of Sclerostin binding by Acid Green 25. The LRP5-ALP was bound to sclerostin in the presence of the indicated concentration of Acid Green 25 used to inhibit this interaction.
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Product Details
Alternative Name: | Sclerostin-LRP binding kit, SOST |
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Assay Time: | 2.5 hours |
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Applications: | FUNC, Chemiluminescence, HTS
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Species reactivity: | Human
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Handling: | Upon receipt, the kit should be stored at -20° C until kit is used. Upon first use of kit, PBS, Wash Buffer, and ALP Substrate may be stored at 4°C. Avoid freeze/thaw cycles. |
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Shipping: | Blue Ice |
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Long Term Storage: | -20°C |
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Contents: | Sclerostin microtiter plate, LRP5-ALP fusion protein concentrate, ALP concentrate (negative control), PBS solution, Blocking agent, wash buffer, acid green 25, ALP substrate reagent, plate sealer |
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Scientific Background: | Sclerostin is encoded by the SOST gene and serves as a negative regulator of bone formation. Wnt ligands bind to Frizzled (Fz) and LRP5/6 receptors to trigger the canonical Wnt signaling cascade, which has been reported to play an important role in bone development. Sclerostin can bind to LRP5/6 to inhibit the canonical Wnt signaling. Antibodies against Sclerostin can block Sclerostin-mediated Wnt inhibition in cell based assays and increase bone mineral density in animal models. Importantly clinical trial studies demonstrate that antibodies against Sclerostin can also increase human bone mineral density. Consequently, Sclerostin is thought to be a superior therapeutic target for osteoporosis treatment over current implemented methods such as treatment with bisphosphonate derivatives. |
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Technical Info/Product Notes: | Application Notes: Detecting LEADING LIGHT® Sclerostin-LRP Screening System using the CLARIOstar® and LVF-Monochromator |
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UniProt ID: | Q9BQB4 |
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Regulatory Status: | RUO - Research Use Only |
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Product Literature References
Methylphenidate enhances neuronal differentiation and reduces proliferation concomitant to activation of Wnt signal transduction pathways: E. Grünblatt, et al.; Transl. Psychiatry
8, 51 (2018),
Abstract;
Full Text
Structural and functional insights into sclerostin-glycosaminoglycan interactions in bone: J. Salbach-Hirsch, et al.; Biomaterials
67, 335 (2015),
Abstract;
WNT1-induced secreted protein-1 (WISP1), a novel regulator of bone turnover and Wnt signaling: A. Maeda, et al.; J. Biol. Chem.
290, 14004 (2015),
Abstract;
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