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SAG

Catalog No. T1779   CAS 912545-86-9
Synonyms: Smoothened Agonist (SAG) HCl, Smoothened Agonist

SAG (Smoothened Agonist) is a potent Smo receptor agonist and activates the Hedgehog signaling pathway.

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SAG Chemical Structure
SAG, CAS 912545-86-9
Pack Size Availability Price/USD Quantity
1 mg In stock $ 47.00
2 mg In stock $ 65.00
5 mg In stock $ 97.00
10 mg In stock $ 155.00
25 mg In stock $ 288.00
50 mg In stock $ 396.00
100 mg In stock $ 587.00
200 mg In stock $ 838.00
500 mg In stock $ 1,250.00
1 mL * 10 mM (in DMSO) In stock $ 98.00
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Purity: 98.88%
Purity: 98.86%
Purity: 98.16%
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Biological Description
Chemical Properties
Storage & Solubility Information
Description SAG (Smoothened Agonist) is a potent Smo receptor agonist and activates the Hedgehog signaling pathway.
Targets&IC50 Smoothened:3 nM(EC50)
In vitro SAG regulates Smo activity by binding directly to the Smo heptahelical bundle. [1] SAG induces Smo-dependent signaling through Gli in a GRK2-dependent way. [2] SAG also (1 nM) induces proliferation of neuronal and glial precursors without affecting the differentiation pattern of newly produced cells. [3]
In vivo In the adult rat hippocampus, the intracerebroventricular administration of SAG (2.5 nM) significantly increases the number of newly generated cells and extends survival of hippocampal cells. [3] In mice, SAG (20 μg/g, i.p.) effectively prevents GC-induced neonatal cerebellar developmental abnormalities. [4]
Kinase Assay In vitro Kinase Assays: Kinase assays for CDK1, CDK2 and GSK3-β are all carried out in a radiometric filter binding format. Assays for CDK5 are in DELFIA format and for CDKs 4 and 6 in ELISA format. For CDKs 1 and 2, the relevant CDK and 0.12 μg/mL Histone H1 are incubated in 20 mM MOPS, pH 7.2, 25 mM β-glycerophosphate, 5 mM EDTA, 15 mM MgCl2, 1 mM sodium orthovanadate, 1 mM DTT, 0.1 mg/mL BSA, 45 μM ATP (0.78 Ci/mmol) and different concentrations of AT7519 for 2 or 4 hours respectively. For GSK3-β, the relevant enzyme and 5 μM glycogen synthase peptide 2 along with 10 mM MOPS pH 7.0, 0.1 mg/mL BSA, 0.001% Brij-35, 0.5% glycerol, 0.2 mM EDTA, 10 mM MgCl2, 0.01% β-mercaptoethanol, 15 μM ATP (2.31 Ci/mmol) and different concentrations of AT7519 are incubated for 3 hours. Assay reactions are stopped by adding an excess of orthophosphoric acid and filtered using Millipore MAPH filter plates. The plates are then washed, scintillant added and radioactivity measured by scintillation counting on a Packard TopCount. For CDK5, CDK5/p35 and 1 μM of a biotinylated Histone H1 peptide (Biotin-PKTPKKAKKL) are incubated in 25 mM Tris-HCl, pH 7.5, 2.5 mM MgCl2, 0.025% Brij-35, 0.1 mg/mL BSA, 1 mM DTT, 15 μM ATP and different concentrations of AT7519 for 30 minutes. Assay reactions are stopped using EDTA, transferred to Neutravidin-coated plates and phosphorylated peptide quantified by means of a rabbit phospho-cdk1 substrate polyclonal antibody and DELFIA europium-labelled anti-rabbit IgG secondary antibody using time-resolved fluorescence at λex=335 nM, λem=620 nM. For CDK 4 and 6 assays, plates are coated with GST- pRb769-921 and blocked with Superblock. CDK4 or 6 is incubated with 15 mM MgCl2, 50 mM HEPES, pH 7.4, 1 mM DTT, 1 mM EGTA, pH 8.0, 0.02% Triton X-100, 2.5% DMSO and different concentrations of AT7519; the reaction is initiated by addition of ATP. After 30 minutes, reactions are stopped by the addition of 0.5 M EDTA pH 8.0.Plates are then washed and incubated for one hour with the primary antibody (anti- p-Rb Serine 780) diluted in Superblock followed by secondary antibody (alkaline phosphatase linked anti-rabbit) for a further hour. Plates are developed using the Attophos system and fluorescence read on a Spectramax Gemini plate reader at excitation 450 nm and emission 580 nm. In all cases, IC50 values are calculated from replicate curves, using GraphPad Prism software.
Synonyms Smoothened Agonist (SAG) HCl, Smoothened Agonist
Molecular Weight 490.06
Formula C28H28ClN3OS
CAS No. 912545-86-9

Storage

Powder: -20°C for 3 years | In solvent: -80°C for 1 year

Solubility Information

DMSO: 71 mg/mL (134.84 mM), warmed

Ethanol: 40 mg/mL (75.97 mM), warmed

TargetMolReferences and Literature

1. Chen JK, et al. Proc Natl Acad Sci U S A. 2002, 99(22), 14071-14076. 2. Meloni AR, et al. Mol Cell Biol. 2006, 26(20), 7550-760. 3. Bragina O, et al. Neurosci Lett. 2010, 482(2), 81-85. 4. Heine VM, et al. Sci Transl Med. 2011, 3(105), 105ra104. 5. Schenke M, Schjeide B M, Püschel G P, et al. Analysis of Motor Neurons Differentiated from Human Induced Pluripotent Stem Cells for the Use in Cell-Based Botulinum Neurotoxin Activity Assays[J]. Toxins. 2020, 12(5): 276. 6. Schenke M. DOCTOR OF PHILOSOPHY (PhD)[D]. University of Veterinary Medicine Hannover. 2020

TargetMolCitations

1. Schenke M, Schjeide B M, Püschel G P, et al. Analysis of Motor Neurons Differentiated from Human Induced Pluripotent Stem Cells for the Use in Cell-Based Botulinum Neurotoxin Activity Assays. Toxins. 2020, 12(5): 276

Related compound libraries

This product is contained In the following compound libraries:
Osteogenesis Compound Library GPCR Compound Library Bioactive Compounds Library Max Anti-Cancer Compound Library Anti-Obesity Compound Library Stem Cell Differentiation Compound Library Anti-Prostate Cancer Compound Library Reprogramming Compound Library Bioactive Compound Library Neuronal Differentiation Compound Library

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Keywords

SAG 912545-86-9 GPCR/G Protein Stem Cells Hedgehog/Smoothened Smo Smoothened Agonist (SAG) HCl inhibit Smoothened Inhibitor Smoothened Agonist inhibitor

 

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