In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
SAR405 is a selective ATP-competitive inhibitor of Vps34 with a Kd value of 1.5 nM.
Vps34 is a phosphoinositide 3-kinase (PI3K) class III isoform that plays important role in autophagy. Vps34 was originally described in yeast to be involved in vesicle trafficking. SAR405 is a first-in-class catalytic Vps34 inhibitor and represents a unique pharmacological tool to investigate the biology around this protein. This compound has an exquisite protein and lipid kinase selectivity profile that is explained by its unique binding mode and molecular interactions within the ATP binding cleft of human Vps34. This is the first potent and specific Vps34 inhibitor described so far. Inhibition of Vps34 kinase activity by SAR405 affects both late endosome-lysosome compartments and prevents autophagy.
Using SAR405, inhibition of Vps34 did not affect early events of endocytosis but resulted in an accumulation of swollen late endosome-lysosomes. A defect of cathepsin D maturation was also identified upon treatment with SAR405, indicating that the function of lysosomes is impaired. This result is in agreement with a previous report using Vps34 siRNA, which affects vesicle trafficking from late endosomes to lysosomes.
SAR405 had an IC50 of 1 nM in the phosphorylation of a PtdIns substrate by human recombinant Vps34 enzyme. This relocalization of the GFP-FYVE indicated that SAR405 inhibits PtdIns3P formation. As SAR405 was found to not be active up to 10 μM on class I and class II PI3Ks as well as on mTOR. Binding of SAR405 to the ATP site of protein and lipid kinases from the lysate of Jurkat cells was measured after incubation at 1μM, which is more than 600-fold the KD for recombinant Vps34. SAR405 did not affect the Akt phosphorylation in the PC3 cell line at concentrations up to 10 μM.
. Ronan B, Flamand O1, Vescovi L et al. A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy. Nat Chem Biol. 2014 Dec;10(12):1013-9.
- 1. Heeseon An, Alban Ordureau, et al. "Systematic quantitative analysis of ribosome inventory during nutrient stress." Nature. 2020 Jul;583(7815):303-309.nbsp;PMID:32612236
- 2. Luciani A, Schumann A, et al. "Impaired mitophagy links mitochondrial disease to epithelial stress in methylmalonyl-CoA mutase deficiency." Nat Commun. 2020;11(1):970. Published 2020 Feb 20.nbsp;PMID:32080200
- 3. Scotto Rosato A, Montefusco S, et al. "TRPML1 links lysosomal calcium to autophagosome biogenesis through the activation of the CaMKKβ/VPS34 pathway." Nat Commun. 2019 Dec 10;10(1):5630.nbsp;PMID:31822666
- 4. An H, Ordureau A, et al. "TEX264 Is an Endoplasmic Reticulum-Resident ATG8-Interacting Protein Critical for ER Remodeling during Nutrient Stress." Mol Cell. 2019 Jun 6;74(5):891-908.e10.nbsp;PMID:31006537
- 5. Yuan NN, Cai CZ, et al. "Canthin-6-One Accelerates Alpha-Synuclein Degradation by Enhancing UPS Activity: Drug Target Identification by CRISPR-Cas9 Whole Genome-Wide Screening Technology." Front Pharmacol. 2019 Jan 28;10:16.nbsp;PMID:30745870
- 6. Zhang M, Liu F, et al. "The MTOR signaling pathway regulates macrophage differentiation from mouse myeloid progenitors by inhibiting autophagy." Autophagy. 2019 Feb 27:1-13. PMID:30724690
- 7. Cui-ZanCai, He-FengZhou, et al. "Natural alkaloid harmine promotes degradation of Alpha-synuclein via PKA-mediated ubiquitin-proteasome system activation." Phytomedicine. Available online 30 January 2019, 152842.
- 8. Jacob M New. "Autophagy in Head and Neck Cancer Associated Fibroblasts: Biology and Therapy." University of Kansas.2018.
- 9. Filippakis H, Belaid A, et al. "Vps34-mediated macropinocytosis in Tuberous Sclerosis Complex 2-deficient cells supports tumorigenesis." Sci Rep. 2018 Sep 21;8(1):14161. PMID:30242175
- 10. Luhr M, Szalai P, Engedal N. "The Lactate Dehydrogenase Sequestration Assay - A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells." J Vis Exp. 2018 Jul 27;(137). PMID:30102280
- 11. Wenzel EM, Schultz SW, et al. "Concerted ESCRT and clathrin recruitment waves define the timing and morphology of intraluminal vesicle formation." Nat Commun. 2018 Jul 26;9(1):2932. PMID:30050131
- 12. McGuire CM, Forgac M. "Glucose Starvation Increases V-ATPase Assembly and Activity in Mammalian Cells through AMP Kinase and Phosphatidylinositide 3-Kinase/Akt Signaling." J Biol Chem. 2018 Mar 14. pii: jbc.RA117.001327. PMID:29540478
- 13. Festa BP, Chen Z, et al. "Impaired autophagy bridges lysosomal storage disease and epithelial dysfunction in the kidney. "Nat Commun.2018 Jan 11;9(1):161. PMID:29323117
- 14. An H, Harper JW. "Systematic analysis of ribophagy in human cells reveals bystander flux during selective autophagy." Nat Cell Biol. 2017 Dec 11. PMID:29230017
- 15. Follo C, Cheng Y, et al. "Inhibition of autophagy initiation potentiates chemosensitivity in mesothelioma." Mol Carcinog. 2018 Mar;57(3):319-332. PMID:29073722
- 16. Heydt Q, Larrue C, et al. "Oncogenic FLT3-ITD supports autophagy via ATF4 in acute myeloid leukemia." Oncogene. 2018 Feb 8;37(6):787-797. PMID:29059168
- 17. Hong Z, Pedersen NM, et al. "PtdIns3P controls mTORC1 signaling through lysosomal positioning. J Cell Biol." 2017 Oct 13. pii: jcb.201611073. PMID:29030394
- 18. New J, Arnold L, et al. "Secretory Autophagy in Cancer-Associated Fibroblasts Promotes Head and Neck Cancer Progression and Offers a Novel Therapeutic Target." Cancer Res. 2017 Dec 1;77(23):6679-6691. PMID:28972076
- 19. Bartolomeo R, Cinque L, et al. "mTORC1 hyperactivation arrests bone growth in lysosomal storage disorders by suppressing autophagy. J Clin Invest." 2017 Oct 2;127(10):3717-3729. PMID:28872463
- 20. Tan HWS, Sim AYL, Long YC. "Glutamine metabolism regulates autophagy-dependent mTORC1 reactivation during amino acid starvation." Nat Commun. 2017 Aug 24;8(1):338. PMID:28835610
- 21. Ge L, Zhang M, et al. "Remodeling of ER-exit sites initiates a membrane supply pathway for autophagosome biogenesis." EMBO Rep. 2017 Sep;18(9):1586-1603. PMID:28754694
- 22. Vdovikova S, Luhr M, et al. "A Novel Role of Listeria monocytogenes Membrane Vesicles in Inhibition of Autophagy and Cell Death." Front Cell Infect Microbiol. 2017 May 3;7:154. PMID:28516064
- 23. Filippakis H, Alesi N, et al. "Lysosomal regulation of cholesterol homeostasis in tuberous sclerosis complex is mediated via NPC1 and LDL-R." Oncotarget. 2017 Jun 13;8(24):38099-38112. PMID:28498820
- 24. Katheder NS, Khezri R, et al."Microenvironmental autophagy promotes tumour growth." Nature. 2017 Jan 19;541(7637):417-420. PMID:28077876
- 25. Uchida Y, Rutaganira FU, et al. "Endosomal Phosphatidylinositol 3-Kinase Is Essential for Canonical GPCR Signaling." Mol Pharmacol. 2017 Jan;91(1):65-73. PMID:27821547
|Storage||Store at -20°C|
|Solubility||≥22.19 mg/mL in DMSO; insoluble in H2O; ≥32.25 mg/mL in EtOH with ultrasonic|
|Canonical SMILES||C[[email protected]]1N(C(N=C2N3CC[[email protected]@H](C(F)(F)F)N2CC4=CN=CC(Cl)=C4)=CC3=O)CCOC1|
|Shipping Condition||Evaluation sample solution: ship with blue ice. All other available sizes: ship with RT, or blue ice upon request.|
|General tips||For obtaining a higher solubility, please warm the tube at 37°C and shake it in the ultrasonic bath for a while. Stock solution can be stored below -20°C for several months.|
|Kinase experiment :|
Vps34 cellular assay
The activity of SAR405 was evaluated on a dedicated Vps34 cellular assay. Using a GFP-FYVE–transfected HeLa cell line, treatment with SAR405 triggered the relocalization of GFP-FYVE inside the cell without affecting GFP intensity. Binding of SAR405 to the ATP site of protein and lipid kinases from the lysate of Jurkat cells was measured after incubation at 1 μM, which is more than 600-fold the KD for recombinant Vps34. Results confirmed very potent binding to endogenous Vps34 (>94% inhibition).
|Cell experiment :|
GFP-LCLC3 HeLa cells ; GFP-LCLC3 H1299 cells
Soluble in DMSO > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.
16 h-24 h
In RKO cells, 24-h SAR405 treatment leads to a dose-dependent accumulation of p62 protein and 16-h of SAR405 treatment shows a significant decrease of mature cathepsin D. SAR405 also completely inhibits the formation of autophagosomes in GFP-LCLC3 HeLa cells. In addition, SAR405 prevents autophagy induced by mTOR inhibitor and synergizes with everolimus in GFP-LCLC3 H1299 cells,
1. Ronan B, Flamand O1, Vescovi L et al. A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy. Nat Chem Biol. 2014 Dec;10(12):1013-9.
Quality Control & MSDS
- View current batch:
Purity = 98.74%
- COA (Certificate Of Analysis)
- MS (Mass Spectrometry)
- MSDS (Material Safety Data Sheet)
Purity = 98.07%
- COA (Certificate Of Analysis)
- MS (Mass Spectrometry)
- MSDS (Material Safety Data Sheet)