Leupeptin hemisulfate
Based on 62 publication(s) in Google Scholar
Leupeptin hemisulfate is a broad-spectrum, membrane-permeable protease inhibitor. Leupeptin hemisulfate potently inhibits serine, cysteine and threonine proteases. Leupeptin hemisulfate inhibits Mpro (the main protease of SARS-CoV-2) and also has anti-inflammatory activity.
For research use only. We do not sell to patients.
- Purity: 99.39%
- CAS No.: 103476-89-7
- Formula: C20H38N6O4.1/2H2SO4
- Molecular Weight:475.59
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Storage:
-20°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Publications Citing Use of MedChemExpress (MCE) Leupeptin hemisulfate
More- Signal Transduct Target Ther. 2025 Dec 15;10(1):406. [Abstract]
- Nature. 2024 Aug;632(8025):686-694. [Abstract]
- Nature. 2024 Aug;632(8026):930-937. [Abstract]
- Nature. 2023 Jun;618(7966):799-807. [Abstract]
- Cell. 2025 Nov 26;188(24):6861-6872.e14. [Abstract]
- Nat Biotechnol. 2025 Sep 10. [Abstract]
- J Hematol Oncol. 2024 Sep 2;17(1):78. [Abstract]
- Cell Res. 2025 May;35(5):385-388. [Abstract]
- Natl Sci Rev. 2021 Feb 10;8(7):nwab024. [Abstract]
- Nat Commun. 2025 Nov 19;16(1):10181. [Abstract]
- Nat Commun. 2025 Jul 16;16(1):6551. [Abstract]
- Nat Commun. 2025 Apr 2;16(1):3153. [Abstract]
- Nat Commun. 2025 Mar 11;16(1):2412. [Abstract]
- Nat Commun. 2019 Jan 24;10(1):411. [Abstract]
- Cell Death Differ. 2023 Jan;30(1):137-151. [Abstract]
- Autophagy. 2025 Aug 6:1-17. [Abstract]
- Autophagy. 2025 Feb;21(2):352-373. [Abstract]
- Autophagy. 2024 Oct;20(10):2221-2237. [Abstract]
- Autophagy. 2021 Aug;17(8):1873-1888. [Abstract]
- Autophagy. 2021 Jul;17(7):1592-1613. [Abstract]
- Adv Sci (Weinh). 2025 Jul 15:e03486. [Abstract]
- J Clin Invest. 2022 Mar 1;132(5):e152170. [Abstract]
- Neuro Oncol. 2023 Jan 5;25(1):82-96. [Abstract]
- Sci Adv. 2026 Jan 30;12(5):eadz8234. [Abstract]
- Carbohydr Polym. 2023 Oct 1:317:121108. [Abstract]
- Sci Adv. 2021 Jan 1;7(1):eabe1340. [Abstract]
- Metabolism. 2026 May:178:156553. [Abstract]
- Redox Biol. 2024 May:71:103100. [Abstract]
- Redox Biol. 2024 Apr:70:103064. [Abstract]
- Pharmacol Res. 2024 Jun 28:107252. [Abstract]
- Mol Biomed. 2023 Nov 17;4(1):42. [Abstract]
- Cancer Lett. 2022 Jun 1:535:215629. [Abstract]
- Cell Death Dis. 2022 Oct 12;13(10):865. [Abstract]
- Int J Biol Macromol. 2023 Aug 30:247:125670. [Abstract]
- Br J Pharmacol. 2023 Aug;180(15):1930-1948. [Abstract]
- Emerg Microbes Infect. 2023 Dec;12(1):2207688. [Abstract]
- Cell Rep. 2025 Apr 2;44(4):115489. [Abstract]
- Cell Rep. 2021 Nov 2;37(5):109931. [Abstract]
- Neurobiol Dis. 2025 Oct 15:215:107090. [Abstract]
- Biochem Pharmacol. 2025 Apr 23:237:116955. [Abstract]
- Biochem Pharmacol. 2024 Dec 19:116720. [Abstract]
- Biochem Pharmacol. 2022 Dec:206:115339. [Abstract]
- Int J Oncol. 2019 Jul;55(1):331-339. [Abstract]
- mBio. 2024 Jun 28:e0053224. [Abstract]
- Structure. 2025 Dec 4;33(12):2049-2057.e5. [Abstract]
- Antiviral Res. 2020 Oct;182:104922. [Abstract]
- Sci Rep. 2022 Jul 16;12(1):12197. [Abstract]
- Oncol Rep. 2021 Dec;46(6):253. [Abstract]
- J Biol Chem. 2021 Jan-Jun:296:100616. [Abstract]
- Biochim Biophys Acta Mol Cell Res. 2023 Jan 24;1870(3):119433. [Abstract]
- Toxicol Appl Pharmacol. 2023 Jul 15:471:116568. [Abstract]
- Chem Asian J. 2024 Sep 2:e202400824. [Abstract]
- Biomed Eng Online. 2026 Apr 13;25(1):74. [Abstract]
- J Inorg Biochem. 2025 Nov:272:113006. [Abstract]
- Environ Toxicol. 2024 Mar;39(3):1505-1520. [Abstract]
- Biochem Biophys Res Commun. 2025 Apr 21:766:151870. [Abstract]
- STAR Protoc. 2025 Jul 18;6(3):103963. [Abstract]
- bioRxiv. 2026 Jan 26:2026.01.23.701155. [Abstract]
- Complutense University of Madrid. 2025.
- bioRxiv. 2024 November 14.
- bioRxiv. 2023 Nov 5.
- bioRxiv. 2020 Jun.
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All Cathepsin Isoforms
More
Biological Activity
Cathepsin B, Cathepsin H, Cathepsin L, Ser/Thr Protease, Mpro[1][2][3].
Leupeptin hemisulfate (0.06-200 µM; 72 h) significantly decreases copy numbers of SARS-CoV-2 viral RNA (vRNA) in Vero cells[1]
Leupeptin hemisulfate inhibits RNA levels of SARS-CoV-2 in Vero cells, with an EC50 value of 42.34 µM[1].
Leupeptin hemisulfate has some inhibitory activity against Mpro, with an IC50 value of 127.2 µM[1].
Leupeptin hemisulfate againsts human coronavirus strain 229E with an IC50 value of 0.4 µg/mL (about 1 µM)[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:Vero cells
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Concentration:0.06-200 µM
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Incubation Time:72 h
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Result:Significantly decreased copy numbers of SARS-CoV-2 viral RNA (vRNA).
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:C57BL/6NCrl male mice (6-8 weeks old, 20-25 g)[1]
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Dosage:0, 9, 18, 36 mg/kg
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Administration:Intraperitoneal injection; single
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Result:Promoted the accumulation of LC3b-II in mouse liver.
Chemical Information
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CAS No. 103476-89-7
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Appearance Solid
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Molecular Weight 475.59
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Formula C20H38N6O4.1/2H2SO4
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Color White to off-white
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SMILES
O=C[C@@H](NC([C@@H](NC([C@@H](NC(C)=O)CC(C)C)=O)CC(C)C)=O)CCCNC(N)=N.[0.5H2SO4]
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Sequence Shortening
Ac-LLR-CHO
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Structure Classification
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Initial Source
Microbial Metabolite
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
-20°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Publications (62)
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Journal Impact Factor
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Most Recent
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Signal Transduct Target Ther
Selective depletion of tumor-associated SAMHD1 enhances chemotherapeutic efficacy and antitumor immune responses. [Abstract]2025 Dec 15;10(1):406. PMID: 41392286 -
Nature
2024 Aug;632(8025):686-694. PMID: 39112701 -
Nature
2024 Aug;632(8026):930-937. PMID: 39085602 -
Nature
2023 Jun;618(7966):799-807. PMID: 37316670 -
Cell
2025 Nov 26;188(24):6861-6872.e14. PMID: 41138730 -
Nat Biotechnol
A rapid imaging-based screen for induced-proximity degraders identifies a potent degrader of oncoprotein SKP2. [Abstract]2025 Sep 10. PMID: 40931108 -
J Hematol Oncol
METTL16-SENP3-LTF axis confers ferroptosis resistance and facilitates tumorigenesis in hepatocellular carcinoma. [Abstract]2024 Sep 2;17(1):78. PMID: 39218945
Leupeptin hemisulfate purchased from MedChemExpress. Usage Cited in: J Hematol Oncol. 2024 Sep 2;17(1):78. [Abstract]
The LTF expression was detected under MG132 (10 μM) or Leupeptin (100 μM) treatment.
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Cell Res
Structural basis of augmenting taurine uptake by the taurine transporter in alleviating cellular senescence. [Abstract]2025 May;35(5):385-388. PMID: 40108449 -
Natl Sci Rev
SNX14 deficiency-induced defective axonal mitochondrial transport in Purkinje cells underlies cerebellar ataxia and can be reversed by valproate. [Abstract]2021 Feb 10;8(7):nwab024. PMID: 34691693 -
Nat Commun
PRDX1 promotes testosterone synthesis and attenuates aging via redox regulation of ATG4B to modulate lipophagy. [Abstract]2025 Nov 19;16(1):10181. PMID: 41261096 -
Nat Commun
2025 Jul 16;16(1):6551. PMID: 40670375 -
Nat Commun
Molecular insights into the α6β4 nicotinic acetylcholine receptor function and ligand recognition. [Abstract]2025 Apr 2;16(1):3153. PMID: 40175361 -
Nat Commun
2025 Mar 11;16(1):2412. PMID: 40069141 -
Nat Commun
Usp7 regulates Hippo pathway through deubiquitinating the transcriptional coactivator Yorkie. [Abstract]2019 Jan 24;10(1):411. PMID: 30679505 -
Cell Death Differ
SOCS2-enhanced ubiquitination of SLC7A11 promotes ferroptosis and radiosensitization in hepatocellular carcinoma. [Abstract]2023 Jan;30(1):137-151. PMID: 35995846
Leupeptin hemisulfate purchased from MedChemExpress. Usage Cited in: Cell Death Differ. 2023 Jan;30(1):137-151. [Abstract]
Western blot analysis of SLC7A11, SOCS2 and tubulin proteins in SK-Hep-1 and HepG2 cells at 4 h after 4 Gy IR. Leupeptin (50 μM) were added before IR.
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Autophagy
Dysfunctional autophagy triggers STING1 activation to exacerbate cartilage degeneration in obesity-associated osteoarthritis. [Abstract]2025 Aug 6:1-17. PMID: 40728163 -
Autophagy
Impaired degradation of PLCG1 by chaperone-mediated autophagy promotes cellular senescence and intervertebral disc degeneration. [Abstract]2025 Feb;21(2):352-373. PMID: 39212196
Leupeptin hemisulfate purchased from MedChemExpress. Usage Cited in: Autophagy. 2025 Feb;21(2):352-373. [Abstract]
Immunoblotting showing the level of PLCG1 in NPC after treatment with Leu (10 μM), NH4Cl (20 mM), or their combination for 12 h. Right panels represent normalized (/NC) band intensities of interest.
Leupeptin hemisulfate purchased from MedChemExpress. Usage Cited in: Autophagy. 2025 Feb;21(2):352-373. [Abstract]
Western blot showing PLCG1 levels in NPC treated with a combination of Leu (10 μM) and NH4Cl (N/L) for 3, 6, and 9 h. Right panels represent normalized (/NC) band intensities of interest.
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Autophagy
PLK2-mediated phosphorylation of SQSTM1 S349 promotes aggregation of polyubiquitinated proteins upon proteasomal dysfunction. [Abstract]2024 Oct;20(10):2221-2237. PMID: 39316746 -
Autophagy
Mice deficient in UXT exhibit retinitis pigmentosa-like features via aberrant autophagy activation. [Abstract]2021 Aug;17(8):1873-1888. PMID: 32744119
Leupeptin hemisulfate purchased from MedChemExpress. Usage Cited in: Autophagy. 2021 Aug;17(8):1873-1888. [Abstract]
661 W cells were transfected with the indicated siRNA for 24 h. Then, 661 W cells were treated with 10 μg/ml Leupeptin for 24 h. Cell lysates were collected for western blot analysis of LC3B and GAPDH.
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Autophagy
Discovery of a potent SCAP degrader that ameliorates HFD-induced obesity, hyperlipidemia and insulin resistance via an autophagy-independent lysosomal pathway. [Abstract]2021 Jul;17(7):1592-1613. PMID: 32432943 -
Adv Sci (Weinh)
LncDARS-AS1 Regulates ATP1A1 Stability and Enhances Na+/K+ ATPase Activity to Promote Osteosarcoma Metastasis. [Abstract]2025 Jul 15:e03486. PMID: 40665639 -
J Clin Invest
USP25 inhibition ameliorates Alzheimer's pathology through the regulation of APP processing and Aβ generation. [Abstract]2022 Mar 1;132(5):e152170. PMID: 35229730 -
Neuro Oncol
2023 Jan 5;25(1):82-96. PMID: 35727735 -
Sci Adv
Structural insight into the glucose-6-phosphate transport by G6PT1 and inhibition mechanism of CGA. [Abstract]2026 Jan 30;12(5):eadz8234. PMID: 41616054 -
Carbohydr Polym
Metabolic degradation of polysaccharides from Lentinus edodes by Kupffer cells via the Dectin-1/Syk signaling pathway. [Abstract]2023 Oct 1:317:121108. PMID: 37364942 -
Sci Adv
Trisomy 21-induced dysregulation of microglial homeostasis in Alzheimer's brains is mediated by USP25. [Abstract]2021 Jan 1;7(1):eabe1340. PMID: 33523861 -
Metabolism
PFKFB3 nuclear translocation improves diabetic retinopathy by attenuating endothelial cell senescence through inhibition of USP7-p53 axis. [Abstract]2026 May:178:156553. PMID: 41655956 -
Redox Biol
IL-13 facilitates ferroptotic death in asthmatic epithelial cells via SOCS1-mediated ubiquitinated degradation of SLC7A11. [Abstract]2024 May:71:103100. PMID: 38484644 -
Redox Biol
SELENOK-dependent CD36 palmitoylation regulates microglial functions and Aβ phagocytosis. [Abstract]2024 Apr:70:103064. PMID: 38320455 -
Pharmacol Res
Cyclopeptide RA-V from Rubia yunnanensis restores activity of Adagrasib against colorectal cancer by reducing the expression of Nrf2. [Abstract]2024 Jun 28:107252. PMID: 38945380 -
Mol Biomed
Aloperine targets lysosomes to inhibit late autophagy and induces cell death through apoptosis and paraptosis in glioblastoma. [Abstract]2023 Nov 17;4(1):42. PMID: 37975957 -
Cancer Lett
Rab8A promotes breast cancer progression by increasing surface expression of Tropomyosin-related kinase B. [Abstract]2022 Jun 1:535:215629. PMID: 35278612 -
Cell Death Dis
Mesenchymal stem cells exosomal let-7a-5p improve autophagic flux and alleviate liver injury in acute-on-chronic liver failure by promoting nuclear expression of TFEB. [Abstract]2022 Oct 12;13(10):865. PMID: 36224178 -
Int J Biol Macromol
ADRM1/RPN13 attenuates cartilage extracellular matrix degradation via enhancing UCH37-mediated ALK5 deubiquitination. [Abstract]2023 Aug 30:247:125670. PMID: 37406898 -
Br J Pharmacol
A novel sorbicillinoid compound as a potent anti-inflammation agent through inducing NLRP3 protein degradation. [Abstract]2023 Aug;180(15):1930-1948. PMID: 36788033 -
Emerg Microbes Infect
Porcine deltacoronavirus resists antibody neutralization through cell-to-cell transmission. [Abstract]2023 Dec;12(1):2207688. PMID: 37125733 -
Cell Rep
LAMP2A-mediated neuronal hyperexcitability by enhancing NKAβ1 degradation underlies depression-induced allodynia. [Abstract]2025 Apr 2;44(4):115489. PMID: 40178973 -
Cell Rep
2021 Nov 2;37(5):109931. PMID: 34731621 -
Neurobiol Dis
2025 Oct 15:215:107090. PMID: 40930427 -
Biochem Pharmacol
GSNO induced mitochondrial Cx43 nitrosylation in cardiomyocyte differentiation from mouse ES cells in vitro. [Abstract]2025 Apr 23:237:116955. PMID: 40280246 -
Biochem Pharmacol
E3 ubiquitin ligase ITCH-mediated proteasomal degradation of WBP2 sensitizes breast cancer cells to chemotherapy through restraining AMOTL2/c-JUN axis. [Abstract]2024 Dec 19:116720. PMID: 39709035 -
Biochem Pharmacol
The ubiquitination of CKIP-1 mediated by Src aggravates diabetic renal fibrosis (original article). [Abstract]2022 Dec:206:115339. PMID: 36347273 -
Int J Oncol
Metformin induces TPC-1 cell apoptosis through endoplasmic reticulum stress-associated pathways in vitro and in vivo. [Abstract]2019 Jul;55(1):331-339. PMID: 31180536 -
mBio
PRRSV GP5 inhibits the antivirus effects of chaperone-mediated autophagy by targeting LAMP2A. [Abstract]2024 Jun 28:e0053224. PMID: 38940560 -
Structure
2025 Dec 4;33(12):2049-2057.e5. PMID: 40914153 -
Antiviral Res
ML-SA1, a selective TRPML agonist, inhibits DENV2 and ZIKV by promoting lysosomal acidification and protease activity. [Abstract]2020 Oct;182:104922. PMID: 32858116 -
Sci Rep
Hepatitis C virus NS3/4A inhibitors and other drug-like compounds as covalent binders of SARS-CoV-2 main protease. [Abstract]2022 Jul 16;12(1):12197. PMID: 35842458 -
Oncol Rep
Berberine exerts its antineoplastic effects by reversing the Warburg effect via downregulation of the Akt/mTOR/GLUT1 signaling pathway. [Abstract]2021 Dec;46(6):253. PMID: 34643248 -
J Biol Chem
The adaptor protein GIPC1 stabilizes the scavenger receptor SR-B1 and increases its cholesterol uptake. [Abstract]2021 Jan-Jun:296:100616. PMID: 33811857 -
Biochim Biophys Acta Mol Cell Res
2023 Jan 24;1870(3):119433. PMID: 36706922 -
Toxicol Appl Pharmacol
Inhibition of CDK9 exhibits anticancer activity in hepatocellular carcinoma cells via targeting ribonucleotide reductase. [Abstract]2023 Jul 15:471:116568. PMID: 37245555 -
Chem Asian J
Selective Protein Degradation through Tetrazine Ligation of Genetically Incorporated Unnatural Amino Acids. [Abstract]2024 Sep 2:e202400824. PMID: 39221720 -
Biomed Eng Online
Synergistic enhancement of peripheral nerve regeneration using electrospun polylactic acid conduits with leupeptin and methylprednisolone in rats. [Abstract]2026 Apr 13;25(1):74. PMID: 41975442 -
J Inorg Biochem
Iridium(III) complexes as type I photosensitizers for hypoxic two-photon photodynamic therapy. [Abstract]2025 Nov:272:113006. PMID: 40682924 -
Environ Toxicol
1-Benzylimidazole attenuates the stemness of breast cancer cells through partially targeting CYP4Z1. [Abstract]2024 Mar;39(3):1505-1520. PMID: 37994574 -
Biochem Biophys Res Commun
Structure-based artificial intelligence-aided design of MYC-targeting degradation drugs for cancer therapy. [Abstract]2025 Apr 21:766:151870. PMID: 40288261 -
STAR Protoc
Protocol for reconstituting enzymatic activities for ultra-large histone methyltransferases NSD1 and SETD2 using a baculovirus expression system. [Abstract]2025 Jul 18;6(3):103963. PMID: 40684435 -
bioRxiv
OSTM1 is a ubiquitin E3 ligase that suppresses B-cell malignancy by activating the cAMP/PKA/CREB pathway. [Abstract]2026 Jan 26:2026.01.23.701155. PMID: 41659680 -
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Solvent & Solubility
H2O : 70 mg/mL (147.19 mM; Need ultrasonic)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
For the following dissolution methods, please prepare the working solution directly:
It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: PBS
Solubility: 100 mg/mL (210.27 mM); Clear solution; Need ultrasonic and warming and heat to 60°C
Purity & Documentation
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Data Sheet (276 KB)
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SDS (394 KB)
- English - EN (394 KB)
- Français - FR (394 KB)
- Deutsch - DE (394 KB)
- Norwegian - NO (394 KB)
- Español - ES (394 KB)
- Swedish - SV (394 KB)
- Italian - IT (394 KB)
- Korean - KR (394 KB)
- Portuguese - PT (394 KB)
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Handling Instructions (2659 KB)
References
[1]. Aoyagi T, et al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68. [Content Brief]
[2]. Fu L, et al. Mechanism of Microbial Metabolite Leupeptin in the Treatment of COVID-19 by Traditional Chinese Medicine Herbs. mBio. 2021 Oct 26;12(5):e0222021. [Content Brief]
[3]. Haspel J, et al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. [Content Brief]
[4]. Aoyagi T, et al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68.
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| H2O | 1 mM | 2.1027 mL | 10.5133 mL | 21.0265 mL | 52.5663 mL |
| 5 mM | 0.4205 mL | 2.1027 mL | 4.2053 mL | 10.5133 mL | |
| 10 mM | 0.2103 mL | 1.0513 mL | 2.1027 mL | 5.2566 mL | |
| 15 mM | 0.1402 mL | 0.7009 mL | 1.4018 mL | 3.5044 mL | |
| 20 mM | 0.1051 mL | 0.5257 mL | 1.0513 mL | 2.6283 mL | |
| 25 mM | 0.0841 mL | 0.4205 mL | 0.8411 mL | 2.1027 mL | |
| 30 mM | 0.0701 mL | 0.3504 mL | 0.7009 mL | 1.7522 mL | |
| 40 mM | 0.0526 mL | 0.2628 mL | 0.5257 mL | 1.3142 mL | |
| 50 mM | 0.0421 mL | 0.2103 mL | 0.4205 mL | 1.0513 mL | |
| 60 mM | 0.0350 mL | 0.1752 mL | 0.3504 mL | 0.8761 mL | |
| 80 mM | 0.0263 mL | 0.1314 mL | 0.2628 mL | 0.6571 mL | |
| 100 mM | 0.0210 mL | 0.1051 mL | 0.2103 mL | 0.5257 mL |
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.