MG-132, proteasome inhibitor (ab141003)
Key features and details
- Potent, reversible proteasome inhibitor
- CAS Number: 133407-82-6
- Purity: > 98%
Soluble in DMSO to 100 mM but unstable for prolonged periods. Soluble in ethanol to 100 mM.
- Form / State: Solid
- Source: Synthetic
Overview
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Product name
MG-132, proteasome inhibitor -
Description
Potent, reversible proteasome inhibitor -
Alternative names
- Z-LLL-CHO
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Biological description
Potent, reversible, proteasome inhibitor (Ki = 4 nM). Inhibits NF-κB activation by preventing IκB degradation (IC50 = 3 μM). Anti-cancer properties in vitro and in vivo. Cell-permeable.
Also available as ethanol solution (ab147047).
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Purity
> 98% -
CAS Number
133407-82-6 -
Chemical structure
Properties
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Molecular weight
475.62 -
Molecular formula
C26H41N3O5 -
Sequence
LLL (Modifications: N-terminal benzyloxycarbonyl; C-terminal aldehyde) -
PubChem identifier
462382 -
Storage instructions
Store at -20°C. It is important to note that this is air sensitive and impurities can occur as a result of air oxidation. Store under desiccating conditions. -
Solubility overview
Soluble in DMSO to 100 mM but unstable for prolonged periods. Soluble in ethanol to 100 mM.
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Handling
Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one week. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.
Need more advice on solubility, usage and handling? Please visit our frequently asked questions (FAQ) page for more details.
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SMILES
CC(C)CC(C=O)NC(=O)C(CC(C)C)NC(=O)C(CC(C)C)NC(=O)OCC1=CC=CC=C1 -
Source
Synthetic
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Research areas
Images
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Chemical Structure - MG-132, proteasome inhibitor (ab141003)2D chemical structure image of ab141003, MG-132, proteasome inhibitor
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Western blot - MG-132, proteasome inhibitor (ab141003)All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
Lane 3 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours, then treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 62 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunocytochemistry/ Immunofluorescence - MG-132, proteasome inhibitor (ab141003)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2μM MG-132 (ab141003) for 18 hours or untreated, labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/100 dilution, followed by Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) secondary antibody at 1/200 dilution (green).
Confocal image showing cytoplasmic staining on HeLa cell line. The expression increased after treatment with 2μM MG-132 (ab141003) for 18 hours.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunocytochemistry/ Immunofluorescence - MG-132, proteasome inhibitor (ab141003)ab62352 staining Nrf2 in untreated HeLa cells (top panel) and treated HeLa cells (bottom panel). Cells were treated with 2µM of MG-132 for 18 hours (ab141003). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab62352 at 1µg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2µg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2µg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Flow Cytometry - MG-132, proteasome inhibitor (ab141003)Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2μM MG-132 (ab141003) for 18 hours (red) or untreated (green), labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/500 dilution compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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Immunoprecipitation - MG-132, proteasome inhibitor (ab141003)SQSTM1 / p62 (phospho S349) was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 2μM MG-132 (ab141003) for 18h with ab211324 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab211324 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate, 10 µg (Input).
Lane 2: ab211324 IP in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211324 in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.Blocking and dilution buffer: 5% NFDM/TBST.
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Western blot - MG-132, proteasome inhibitor (ab141003)All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution
Lane 1 : Untreated C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : C6 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 62 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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Western blot - MG-132, proteasome inhibitor (ab141003)All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 62 kDa why is the actual band size different from the predicted?
Exposure time: 4 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Western blot - MG-132, proteasome inhibitor (ab141003)All lanes : Anti-HIF-1 alpha antibody [EPR16897] (ab179483) at 0.163 µg/ml
Lane 1 : Untreated C6 (rat glial tumor glial cell), whole cell lysate
Lane 2 : C6 treated with 400 µM CoCl2 and 200 µM MG-132 (ab141003) for 4 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 110 kDa why is the actual band size different from the predicted?
Exposure time: 26 secondsBlocking and diluting buffer: 5% NFDM/TBST.
The expression of HIF-1 alpha is induced by CoCl2 and maintained by MG-132 (PMID: 15836611).
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Western blot - MG-132, proteasome inhibitor (ab141003)All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/5000 dilution
Lane 1 : Untreated HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg
Lane 2 : HT-1080 treated with 10µM MG-132 (ab141003) for 4 hours, whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Exposure time: 103 secondsBlocking/diluting buffer and concentration: 5% NFDM/TBST.
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Western blot - MG-132, proteasome inhibitor (ab141003)All lanes : Anti-ATF-4 antibody [EPR18111] (ab184909) at 1/1000 dilution
Lane 1 : Untreated HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (control)
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) treated with 5 µM MG-132 (ab141003) and 3 µg/ml tunicamycin (ab120296) for 6 hours whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 50 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 22095285).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
References (23)
ab141003 has been referenced in 23 publications.
- Saha S et al. Leishmania donovani Targets Host Transcription Factor NRF2 To Activate Antioxidant Enzyme HO-1 and Transcriptional Repressor ATF3 for Establishing Infection. Infect Immun 89:e0076420 (2021). PubMed: 33820818
- Feng D et al. Regulation of Wnt/PCP signaling through p97/VCP-KBTBD7-mediated Vangl ubiquitination and endoplasmic reticulum-associated degradation. Sci Adv 7:N/A (2021). PubMed: 33990333
- Kumar Singh R et al. KSHV-encoded vCyclin can modulate HIF1a levels to promote DNA replication in hypoxia. Elife 10:N/A (2021). PubMed: 34279223
- Robu M et al. Methods to Study Intracellular Movement and Localization of the Nucleotide Excision Repair Proteins at the DNA Lesions in Mammalian Cells. Front Cell Dev Biol 8:590242 (2020). PubMed: 33282869
- Chen K et al. Parkin ubiquitinates GATA4 and attenuates the GATA4/GAS1 signaling and detrimental effects on diabetic nephropathy. FASEB J 34:8858-8875 (2020). PubMed: 32436607
