MG-132

MG-132 is an inhibitor of proteasome with IC50 of 100 nM in a cell-free assay, and also inhibits calpain with IC50 of 1.2 μM.

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MG-132 Chemical Structure

MG-132 Chemical Structure
Molecular Weight: 475.62

Validation & Quality Control

Product Use Citation(40)

Customer Product Validation(9)

Quality Control & MSDS

Related Compound Libraries

MG-132 is available in the following compound libraries:

Proteasome Inhibitors with Unique Features

Product Information

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  • Research Area
  • Combination Therapy
    Combination Therapy

Product Description

Biological Activity

Description MG-132 is an inhibitor of proteasome with IC50 of 100 nM in a cell-free assay, and also inhibits calpain with IC50 of 1.2 μM.
Targets Proteasome [1]
(Cell-free assay)
IC50 100 nM
In vitro MG-132 displays >1000 times more activity than ZLLal in inhibiting the ZLLL-MCA-degrading activity of 20S proteasome with IC50 of 100 nM versus 110 μM. MG-132 also inhibits calpain with IC50 of 1.2 μM. MG-132 induces neurite outgrowth in PC12 cells at an optimal concentration of 20 nM, displaying 500 times more potency than ZLLal. [1] MG-132 (10 μM) potently inhibits TNF-α-induced NF-κB activation, interleukin-8 (IL-8) gene transcription, and IL-8 protein release in A549 cells by inhibition of proteasome-mediated IκBα degradation. [2] MG-132 treatment potently induces p53-dependent apoptosis in KIM-2 cells by 26S proteasome inhibition. [3] Unlike BzLLLCOCHO or PS-341, MG-132 treatment results in weak inhibition of the chymotrypsinlike (CT-L) and peptidylglutamyl peptide hydrolysing (PGPH) activities of the 26S proteasome, whereas multiple myeloma cells (U266 and OPM-2) are more sensitive to induction of apoptosis by MG-132 than BzLLLCOCHO. [4] MG-132 (1 μM) sensitizes TRAIL-resistant prostate cancer cells by activating the AP-1 family members c-Fos and c-Jun, which, in turn, repress the antiapoptotic molecule c-FLIP(L). [5] MG-132 significantly enhances the ability of inositol hexakisphosphate (IP6) to reduce cellular metabolic activity in both PC3 and DU145 androgen-independent prostate cancer (AIPCa) cell lines. [6]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity Description
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IFN-gamma-induced RAW264.7NV;HPJVHTnWwY4Tpc44hSXO|YYm=M37TS2ROW09?MWnJcohq[mm2czDubZRzcWNib4jp[IUheHKxZIXjeIlwdiC5aYToJGlEPTBib3[gNE4xPzdizszN
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... Click to View More Cell Line Experimental Data

In vivo Administration of MG-132 effectively rescues the expression levels and plasma membrane localization of dystrophin, β-dystroglycan, α-bdystroglycan, and α-sarcoglycan in skeletal muscle fibers from mdx mice, reduces muscle membrane damage, and ameliorates the histopathological signs of muscular dystrophy. [7] MG-132 treatment significantly reduces immobilization-induced skeletal muscle atrophy in mice, by downregulating the muscle-specific ubiquitin ligases atrogin-1/MAFbx and MuRF-1 mRNA. [8]
Features

Protocol(Only for Reference)

Kinase Assay:

[1]

Measurement of inhibitory activities of MG-132 against 20S proteasome The reaction mixture for the 20S proteasome inhibitory assay contains 0.1 M Tris-acetate, pH 7.0, 20S proteasome, MG-132, and 25 μM substrate dissolved in dimethyl sulfoxide in a final volume of 1 mL. After incuba tion at 37 °C for 15 minutes, the reaction is stopped by the addition of 0.1 mL of 10% SDS and 0.9 mL of 0.1M Tris acetate, pH 9.0. The fluorescence of the reaction products is measured. To determine the IC50 against 20S proteasome, various concentrations of MG-132 are included in the assay mixture.

Cell Assay:

[3]

Cell lines KIM-2, HC11, and ES
Concentrations Dissolved in DMSO, final concentrations ~25 μM
Incubation Time 24, and 48 hours
Method

Cells are exposed to various concentrations of MG-132 for 24, and 48 hours. Supernatant and monolayer cells are harvested by centrifugation and fixed in 70% ethanol in PBS for staining with acridine orange. Equal volumes of cells and acridine orange (5 mg/mL in PBS) are mixed on a microscope slide and examined by fluorescence microscopy. For annexin V analysis, cells are harvested by centrifugation and stained with annexin V and propidium iodide. For cell cycle analysis, cells are rehydrated in PBS at room temperature for 10 minutes, followed by staining with propidium iodide (5 mg/mL). All samples are analyzed using a Coulter Epics XL flow cytometer.

Animal Study:

[7]

Animal Models Male mdx (C57BL/10ScSn DMD mdx) mice
Formulation Dissolved in DMSO, and diluted in PBS
Dosages ~10 μg/kg/day
Administration Injection

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Tsubuki S, et al. J Biochem, 1996, 119(3), 572-576.

[2] Fiedler MA, et al. Am J Respir Cell Mol Biol, 1998, 19(2), 259-268.

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Chemical Information

Download MG-132 SDF
Molecular Weight (MW) 475.62
Formula

C26H41N3O5

CAS No. 133407-82-6
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 95 mg/mL (199.73 mM)
Ethanol 95 mg/mL (199.73 mM)
Water <1 mg/mL (<1 mM)
In vivo 1% DMSO/30% polyethylene glycol/1% Tween 80, pH 4 6 mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name benzyl (S)-4-methyl-1-((S)-4-methyl-1-((S)-4-methyl-1-oxopentan-2-ylamino)-1-oxopentan-2-ylamino)-1-oxopentan-2-ylcarbamate

Customer Product Validation (9)


Click to enlarge
Rating
Source Nat Cell Biol 2015 17(1), 95-103. MG-132 purchased from Selleck
Method Immunoprecipitation
Cell Lines MDA-MB-231 cells
Concentrations 10 uM
Incubation Time 6 h
Results SIAH2 destabilizes LATS2 through proteasome. SIAH2-mediated LATS2 degradation was inhibited by proteasomal inhibitor MG132, but not by lysosomal inhibitor BA-1. The half-life of LATS2 was shortened by SIAH2.

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Source Nucleic Acids Res 2014 42(1), 458-74. MG-132 purchased from Selleck
Method Western blot
Cell Lines HeLa cells
Concentrations 20 uM
Incubation Time 8 h
Results As RMND5A downregulation resulted in a rapid decrease of protein Exportin-5, this effect was partially abolished by the proteasome inhibitor MG132, indicating that RMND5A protect Exportin-5 protein against proteasome degradation.

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Source Nucleic Acids Res 2014 42(1), 458-74. MG-132 purchased from Selleck
Method Immunocytochemistry
Cell Lines HeLa cells
Concentrations 20 uM
Incubation Time 8 h
Results It was surprised to observe that Exportin-5 retention in nucleus when its protein levels were downregulated by RMND5A siRNA or miR-138. The proteasome inhibitor MG132 could recover not only Exportin-5 protein levels but also its normal subcellular localization.

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Source EMBO Mol Med 2013 5, 397-412. MG-132 purchased from Selleck
Method Western Blot
Cell Lines MEF cells
Concentrations 30 uM
Incubation Time 6 h
Results Treatment of MEF with proteasome inhibitor MG132 resulted in similar increase of steady state levels of intracellular ATZ in both GFP- and TFEB- transfected Cells.

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Source Cell Rep 2015 11(9), 1458-73. MG-132 purchased from Selleck
Method Western Blot
Cell Lines A375, Lu1205 cells
Concentrations 5 uM
Incubation Time 10 h
Results MG132 treatment increased JAK1 steady-state levels, masking the effect of RNF125 depletion and partially blocking deregulated JAK1 expression by ectopically expressed RNF125.

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Source Plant Physiol 2012 60(1), 118-34. MG-132 purchased from Selleck
Method Immunoblot analysis
Cell Lines Arabidopsis thaliana
Concentrations 100 uM
Incubation Time 12 h
Results These rates of loss were markedly slower in the lrb1-1 lrb2-1 background. Importantly, phyB breakdown was also substantially attenuated upon exposing wild-type seedlings to the proteasome inhibitor MG132, thus implicating the 26S proteasome in particular. MG132 treatment also slightly stabilized phyB levels in lrb1-1 lrb2-1 seedlings, implying that other factors besides LRB1 and LRB2 might also direct phyB breakdown.

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Source Toxicol Appl Pharmacol 2015 286(2), 135-41. MG-132 purchased from Selleck
Method Immunofluorescence
Cell Lines γ-H2AX-stained cells
Concentrations 100 uM
Incubation Time 3 h
Results G2 cell cycle arrest usually reflects the formation of genomic damage in the preceding S-phase, which activates checkpoint response upon the entry of cells into G2 phase. Furthermore, MG132-treated cells also had larger amounts of γ-H2AX, as evidenced by the increased staining in individual nuclei .

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Source FEBS Lett 2012 586, 3787–3792. MG-132 purchased from Selleck
Method Western Blot
Cell Lines Hela cells
Concentrations
Incubation Time 12 h
Results When GSK-3 (Ser9) degradation was blocked by proteasome inhibitor (MG132), the increased amount of phosphorylated GSK-3 (Ser9) in responding to NOK was more clearly observed in the HeLa-NOK-HA cells (Fig. C).

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Source 2012 Dr. Edison DUKE-NUS Graduate Medical School Singapore.. MG-132 purchased from Selleck
Method Western blot
Cell Lines NIH3T3 cells
Concentrations 25 μM
Incubation Time 1-4 h
Results By adding MG132, a proteasome inhibitor, the degradation of Per2 was dramatically decreased within 4 hours treatment.

Product Use Citation (40)

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