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Leupeptin,Microbial
本产品不向个人销售,仅用作科学研究,不用于任何人体实验及非科研性质的动物实验。
Leupeptin,Microbial图片
包装与价格:
包装价格(元)
10mM (in 1mL DMSO)电议
25mg电议

产品介绍

Leupeptin, Microbial is a broad-spectrum serine and cysteine protease inhibitor.

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 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 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.

文献引用
产品描述

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.