Leupeptin, Microbial (Synonyms: Acetyl-L-leucyl-L-leucyl-L-argininal)

Name: Leupeptin, Microbial
Brand: GlpBio
SKU: GC10027
Availability: InStock
Rating: 5 (30 reviews)

Catalog No.GC10027

A reversible protease inhibitor

Products are for research use only. Not for human use. We do not sell to patients.

Leupeptin, Microbial Chemical Structure

Cas No.: 103476-89-7

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10mM (in 1mL DMSO)	$36.00	In stock
5mg	$23.00	In stock
25mg	$69.00	In stock
50mg	$112.00	In stock

Tel：(909) 407-4943 Email: sales@glpbio.com

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Sample solution is provided at 25 µL, 10mM.

Product has been cited by 1 publications

J Cell Mol Med 25.6 (2021): 2806-2815.PMID:33538080

Description Protocol Chemical Properties Product Documents

Product Documents

Quality Control & SDS

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Purity: >98.00%

Protocol

Leupeptin analysis [1]:
Preparation Method	The concentration of Leupeptin, Microbial was calculated by enzyme inhibition assay using 50 pg of papain and 0.2 mM of N-benzoylL-arginine ethyl ester as the target protease and papain substrate, respectively. Enzyme reactions were carried out at 25 ℃ and pH 6.2.
Applications	Leupeptin, Microbial is a leucine-specific protease and a metalloprotease.
Cell experiment [2]:
Cell lines	MRC-C cell
Preparation Method	To investigate the effect of Leupeptin, Microbial on virus yield, MRC-C cultures in 35 mm dishes were infected by adsorption of virus from 0T ml concentrated suspension of HCV 229E for 1 h at 36 ℃. The cultures were washed twice and overlaid with 2 ml Eagle’s MEM plus 0.2% bovine serum albumin and 20 mm-HEPES buffer pH 7.4. Leupeptin, Microbial at a range of concentrations was added to the virus inoculum, to the maintenance medium and also to maintenance medium used to treat the cultures for 1 h before infection. After 24 h at 36 ℃ in air, the cells were scraped into the medium and disrupted by sonication.
Reaction Conditions	0.4 to l00 µg/ml for 25 h（1 h before infection to 24 h after infection）
Applications	The protease inhibitor Leupeptin, Microbial prevented multiplication of the human coronavirus strain 229E in cultures of MRC-C cells.
Animal experiment [3]:
Animal models	C57BL/6NCrl male mouse
Preparation Method	Mice received i.p. injections of 0.5 ml sterile Phosphate Buffered Saline or 0.5 ml PBS containing 9–40 mg/kg Leupeptin, Microbial hemisulfate.
Dosage form	0, 9, 18 36 and 40 mg/kg Leupeptin, Microbial; Intraperitoneal injection
Applications	Rate of LC3b accumulation after Leupeptin, Microbial treatment was greatest in the liver and lowest in spleen.
References: [1]. Kim IS, Kim YB, et,al. Characterization of the leupeptin-inactivating enzyme from Streptomyces exfoliatus SMF13 which produces leupeptin. Biochem J. 1998 Apr 15;331 (Pt 2)(Pt 2):539-45. doi: 10.1042/bj3310539. PMID: 9531495; PMCID: PMC1219386. [2]. Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. J Gen Virol. 1985 Feb;66 (Pt 2):363-6. doi: 10.1099/0022-1317-66-2-363. PMID: 3968542. [3]. Haspel J, Shaik RS, et,al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. doi: 10.4161/auto.7.6.15100. Epub 2011 Jun 1. PMID: 21460622; PMCID: PMC3127049.

Leupeptin, Microbial is a broad-spectrum serine and cysteine protease inhibitor ^[1]. Leupeptin, Microbial is a reversible protease inhibitor, It acts on bovine trypsin, human plasminase, bovine splenic cathepsin B, and recombinant human caltrypsin with Ki values of 35 nM, 3.4 μM, 6 nM, and 72 nM^[7].

The protease inhibitor Leupeptin, Microbial prevented multiplication of the human coronavirus strain 229E in cultures of MRC-C cells ^[3]. Leupeptin, Microbial-mediated inhibition of trypsin-like proteases maintains substrate mycelium development, whereas proteolytic degradation of Leupeptin, Microbial in stationary phase cultures derepresses the trypsin-like proteases, leading to the digestion of substrate mycelium and promotion of aerial mycelium formation ^[2]. As the calpain inhibitor Leupeptin, Microbial primarily protected hair cells from neomycin^[5]. The expression of hepatitis B surface antigen (HBsAg) from Leupeptin recovered up to 50% of the cell suspension culture ^[6].

Leupeptin, Microbial was well tolerated in mice. Leupeptin, Microbial significantly increased LC3b-II in a dose-dependent way in the total tissue extract and lysosomal enrichment portion (LE portion). At the level of electron microscopy (EM), Leupeptin, Microbial induced the accumulation of electron-dense vesicle structures ^[4].

References:
[1]. Aoyagi T, Miyata S, et,al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68. doi: 10.7164/antibiotics.22.558. PMID: 4243683.
[2]. Kim IS, Kim YB, et,al. Characterization of the leupeptin-inactivating enzyme from Streptomyces exfoliatus SMF13 which produces leupeptin. Biochem J. 1998 Apr 15;331 ( Pt 2)(Pt 2):539-45. doi: 10.1042/bj3310539. PMID: 9531495; PMCID: PMC1219386.
[3]. Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. J Gen Virol. 1985 Feb;66 ( Pt 2):363-6. doi: 10.1099/0022-1317-66-2-363. PMID: 3968542.
[4]. Haspel J, Shaik RS, et,al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. doi: 10.4161/auto.7.6.15100. Epub 2011 Jun 1. PMID: 21460622; PMCID: PMC3127049.
[5]. Aoyagi T, Miyata S, et,al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68. doi: 10.7164/antibiotics.22.558. PMID: 4243683.
[6]. Coffin AB, Williamson KL, et,al. Profiling drug-induced cell death pathways in the zebrafish lateral line. Apoptosis. 2013 Apr;18(4):393-408. doi: 10.1007/s10495-013-0816-8. PMID: 23413197; PMCID: PMC3627356.
[7]. Ganapathi TR, Sunil Kumar GB, et,al. Analysis of the limitations of hepatitis B surface antigen expression in soybean cell suspension cultures. Plant Cell Rep. 2007 Sep;26(9):1575-84. doi: 10.1007/s00299-007-0379-7. Epub 2007 May 30. PMID: 17534624.

Cas No.	103476-89-7	SDF
Synonyms	Acetyl-L-leucyl-L-leucyl-L-argininal
Chemical Name	2-acetamido-N-(1-((5-((diaminomethylene)amino)-1-oxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)-4-methylpentanamide
Canonical SMILES	O=C(C(C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H])([H])N([H])C(C([H])([H])[H])=O)N([H])C(C(N([H])C(C([H])=O)([H])C([H])([H])C([H])([H])C([H])([H])/N=C(N([H])[H])/N([H])[H])=O)([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H]
Formula	C₂₀H₃₈N₆O₄.1/2H₂SO₄	M.Wt	493.6
Solubility	≥ 24.7mg/mL in DMSO， ≥ 20mg/mL in Water	Storage	Store at -20°C
General tips	Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition	Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table

Prepare stock solution
		1 mg	5 mg	10 mg
	1 mM	2.0259 mL	10.1297 mL	20.2593 mL
	5 mM	0.4052 mL	2.0259 mL	4.0519 mL
	10 mM	0.2026 mL	1.013 mL	2.0259 mL

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In vivo Formulation Calculator (Clear solution)

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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.

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Average Rating: 5 ★★★★★ (Based on Reviews and 30 reference(s) in Google Scholar.)

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Leupeptin, Microbial (Synonyms: Acetyl-L-leucyl-L-leucyl-L-argininal)

Customer Reviews

GlpBio Citations

Bioactive Compounds Premium Provider

Product has been cited by 1 publications

Product Documents

Quality Control & SDS

Protocol

Complete Stock Solution Preparation Table

Molarity Calculator

Dilution Calculator

In vivo Formulation Calculator (Clear solution)

Reviews

Leupeptin, Microbial (Synonyms: Acetyl-L-leucyl-L-leucyl-L-argininal)

Customer Reviews

GlpBio Citations

Bioactive Compounds Premium Provider

Product has been cited by 1 publications

Product Documents

Quality Control & SDS

Protocol

Complete Stock Solution Preparation Table

Molarity Calculator

Dilution Calculator

In vivo Formulation Calculator (Clear solution)

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Reviews