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(42)

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 DescriptionPMID
LPS-stimulated RAW264.7 cellsM17PeWZ2dmO2aX;uJGF{e2G7NUHvfYdPOjVizszNMWXEUXNQMW\Jcohq[mm2czDubZRzcWNib4jp[IUheHKxZIXjeIlwdiC5aYToJGlEPTBib3[gNE4yKM7:TR?=MoLzNlQ3QTd2OU[=
HL-60MkXWR5l1d3SxeHnjJGF{e2G7MmPTOFAh|ryPMlewOFghcA>?MoX4SG1UVw>?MkLZTWM2ODxzMDFOwG0>MWeyOFY6PzR7Nh?=
SMMC-7721NFTN[5ZEgXSxdH;4bYMhSXO|YYm=MoXHOFAh|ryPM4X0N|Q5KGh?M{PqTGROW09?NGHHOZFKSzVyPUeuNUDPxE1?NHrQ[I4zPDZ7N{S5Oi=>
A-549Mk\jR5l1d3SxeHnjJGF{e2G7MWm0NEDPxE1?NXjW[oFoPDhiaB?=NWDBZY1uTE2VTx?=MXrJR|UxRDFyIN88US=>NUXTdWlUOjR4OUe0PVY>
MCF-7M3q2[GN6fG:2b4jpZ{BCe3OjeR?=M{D6RVQxKM7:TR?=NWXuWlhEPDhiaB?=NGjpTm9FVVORMVHJR|UxRTdwMzFOwG0>NUT4dGR2OjR4OUe0PVY>
SW-480NUfOXZA6S3m2b4TvfIlkKEG|c3H5NFrMO4Q1OCEQvF2=M2PsUFQ5KGh?NEDq[2tFVVORNGHvO4RKSzVyPUSg{txOMmi0NlQ3QTd2OU[=
NCI-H929M{juUGN6fG:2b4jpZ{BCe3OjeR?=NVn2[4NIOSEQvF2=NFTjTGQ4OiCqM1POPGROW09?M4DuOmlEPTB;MD6xO{DPxE1?MVGyOFYzPTB6OB?=
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MDA-MB-231M{XKcWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7NYC4U2kyOSEQvF2=MYO3NkBpM{TnfWROW09?M4O1SGlEPTB;MD6xPEDPxE1?MV2yOFE2OzJyNh?=
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LPS-stimulated RAW264.7 cellsMn;jSpVv[3Srb36gRZN{[Xl?M{\iTWROW09?NVjWcFc4UW6qaXLpeJMhVkZva3HwdIFDKESQQTDibY5lcW6pMV:xO|QxPzJ5Nx?=
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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)


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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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Rating
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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Rating
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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Rating
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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Rating
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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Rating
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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Rating
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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Rating
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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Rating
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 (42)

Tech Support & FAQs

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

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