Dulbecco’s modified Eagle’s medium (DMEM, Cat# 10313-021), Z150 L-glutamine 100× (Cat# 25030-081), and Trypsin-EDTA (0.05%) phenol red (Cat# 25300062) were from Gibco by Life Technologies, Australia. SERANA fetal bovine serum (FBS) (Cat# FBS-AU-015, batch no. 18030416) was from Fisher Biotechnology, Australia. cOmplete, mini EDTA-free protease inhibitor cocktail (Cat# 11836170001), PhosSTOP Phosphatase Inhibitors (Cat# 4906837001), hydrogen peroxide 30% (w/w) solution (Cat# H1009), AZD2461 (Cat# SML 1858), and MG132 (Z-Leu-Leu-Leu-al, Cat# C2211) were from Sigma-Aldrich.
Z leu leu leu al mg132
Manufactured by Merck Group
Sourced in United States
Z-Leu-Leu-Leu-al (MG132) is a synthetic peptide that functions as a proteasome inhibitor. It blocks the activity of the proteasome, a complex of enzymes responsible for the degradation of proteins within cells. This product is commonly used in research applications to study cellular processes related to protein homeostasis and regulation.
Automatically generated - may contain errors
Lab products found in correlation
Cycloheximide (chx), by Merck Group (5 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Anti flag m2, by Merck Group (2 mentions)
3 methyladenine 3 ma, by Selleck Chemicals (2 mentions)
Mitomycin c, by Merck Group (2 mentions)
Trypsin edta 0.05 phenol red, by Thermo Fisher Scientific (1 mentions)
Phosstop phosphatase inhibitor, by Merck Group (1 mentions)
Azd2461, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
U2os cell, by American Type Culture Collection (1 mentions)
Reporter lysis buffer, by Promega (1 mentions)
Effectene transfection reagent, by Qiagen (1 mentions)
Doxycycline hydrochloride, by Merck Group (1 mentions)
Mccoy s 5a glutamax, by Thermo Fisher Scientific (1 mentions)
6 thioguanine 6 tg, by Merck Group (1 mentions)
Dmem glutamax, by Thermo Fisher Scientific (1 mentions)
Mycoalert mycoplasma detection kit, by Lonza (1 mentions)
Chloroquine diphosphate, by Merck Group (1 mentions)
N acetylglucosamine, by Merck Group (1 mentions)
Wheat germ agglutinin (wga), by Merck Group (1 mentions)
27 protocols using z leu leu leu al mg132
1
Cell Culture Reagents and Inhibitors
2
Cell Line Stress Response Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK, HeLa, and U2OS cells were obtained from ATCC and routinely tested for mycoplasma infection. The cell lines were not authenticated. Cells were incubated at 37 °C and 5% CO2 in high Glucose GlutaMAX DMEM (Thermo Fisher). Heat shock was performed by incubating cells at 43 °C for 1 h. Proteasome inhibition was performed by treating cells with 10 μM of MG132 (Z-Leu-Leu-Leu-al; Sigma) for 8 h. For HeLa and U2OS cell culture replicates, ~200 million cells were cultured. For HEK cell culture replicates, ~300 million cells were cultured.
3
Knock-Down of HERC2 in U2OS Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human fibroblasts and ethical statements were previously described [5 (link)]. U2OS cells were obtained from ATCC. Cells were cultured at 37°C in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 100 units/ml penicillin, 100 μg/ml streptomycin and 2 mM glutamine. Transfection of cells with siRNAs (Non Targeting, NT: UAGCGACUAAACACAUCAA; HERC2, H2: GACUGUAGCCAGAUUGAAA) purchased from GenePharma was carried out using calcium phosphate. Transfected cells were analyzed 72 hours post-transfection. MG132 (Z-Leu-Leu-Leu-al) (Sigma-Aldrich) was added to the cells for 6 hours to a final concentration of 10 μM.
4
Measuring NF-κB and IGFBP1 Promoter Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Approximately 1.0×105 cells were seeded into 6-well plates (AGC Techno Glass, Shizuoka, Japan) 24 h prior to transfection. For the purpose of detecting the relative activity of NF-κB in Huh7 and IHH cells, the cells were co-transfected with 0.3 µg pHBV/pCXN2-HBx and 0.1 µg pNF-κB-luc, respectively in Effectene transfection reagent (Qiagen). The cells were treated for 24 h with 10 ng/ml TNF-α and/or 10 ng/ml IL-1β, with or without 1 µM MG132 (Z-Leu-Leu-Leu-al; Sigma-Aldrich) after transfection. For detecting the relative activity of IGFBP1-promoter in HepG2 cells, the cells were cotransfected with 0.2 µg pCXN2, pCXN2-HBX, pCMVHBV or pHBV and 0.1 µg pIGFBP1-luc in Effectene transfection reagent (Qiagen). At 30 h post-transfection, the cells were lysed with reporter lysis buffer (Promega, Madison, WI, USA) and the luciferase activities were determined using a luminometer (Luminescencer-JNR II AB-2300; ATTO Bio Instrument, Tokyo, Japan) as described previously (23 (link)).
5
Characterization of Human Colon Cancer Cell Lines
6
α-Synuclein Proteinopathy Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Most chemicals in this study, such as phorbol 12-myristate 13-acetate (PMA; Sigma P1585), N-ethylmaleimide (NEM; Sigma E3876), Chx (Sigma C7698), chloroquine diphosphate (Sigma C6628), MG132 Z-Leu-Leu-Leu-al (Sigma C2211), lectin also known as WGA (Sigma L9640), N-acetylglucosamine (Sigma PHR1432), MPP+ (Sigma D048), thioflavin T (T3516), MPTP (M0896), NaF (201154), and Na3VO4 (567540) were purchased from EMD-Sigma. PFF of α-syn was obtained from StressMarq (SPR-324C) and PMSF (Roche 11359061001), BSA (Fisher bioreagent BP9703), and PBS (Invitrogen 10010031) were used as well. Protease inhibitor used was EDTA-free Halt protease inhibitor cocktail (Thermo Scientific 87785).
7
Analyzing Autophagy in 2fTGH Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
2fTGH (human fibrosarcoma), Flag-HA-TG2 2fTGH, GFP-LC3 2fTGH and RFP-GFP-LC3 2fTGH cell lines were cultured in Dulbecco's modified Eagle's medium (Lonza) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 μg/ml streptomycin and 100 units/ml penicillin at 37°C. For generation of the TG2 stably expressing cells (Flag-HA-TG2 2fTGH), 2fTGH cells were transfected with pLPCX-TG2-HA-Flag using CaCl2 method. Autophagy was induced by using 1 μM rapamycin (Rapam, Sigma-Aldrich) or 10 mM 2-deoxy-D-glucose (2-DG, Sigma-Aldrich). For amino acids starvation (Starv), cells were washed twice in the amino-acid-free medium Earle's Balanced Salt Solution (EBSS) (Sigma-Aldrich) and incubated in the same medium for the indicated periods. In order to inhibit autophagy, cells were incubated in full medium with 20 mM ammonium chloride (NH4Cl, Sigma-Aldrich) for the indicated period. For proteasome inhibition, cells were incubated with 5 μM MG132 (Z-Leu-Leu-Leu-al; Sigma Aldrich) for 4 h. To induce mitochondrial damage, cells were incubated in full medium with 15 μM carbonyl cyanide m-chrolorophenyl hydrazine (CCCP, Sigma-Aldrich). TG2 transamidating activity was inhibited by incubating the cells with 40 μM Z-DON (Zedira) for 24 hours, concomitantly with starvation.
8
Doxorubicin-Induced DNA Damage Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A375 melanoma cells (ATCC CRL-1619) were cultured in DMEM supplemented with 10% fetal bovine serum, 100 μg/ml streptomycin and 100 units/ml penicillin, at 37 °C and 5% CO2 in a humidified atmosphere. Mycoplasma contamination was tested in all cell lines. Cells were treated with Doxorubicin (Sigma D-1515) 5 μM for 6, 16, and 24 h to induce DNA damage. TG2 transamidating activity was inhibited incubating the cells with 40 μM Z-DON (Zedira, Darmstadt, Germany) for 16 h. Z-VAD (Invivogen) was used 20 μM for 16 h to inhibit caspases. To block proteasome activity, cells were incubated in the presence of 5 μM MG132 (Z-Leu-Leu-Leu-al, Sigma-Aldrich) for 16 h. For autophagy inhibition cells were incubated with NH4Cl 20 mM for 16 h. A375 were transfected using lipofectamine 2000 (Invitrogen), following the manufactories instructions. 10 nM of oligunocleotides siRNA TGM2 (OriGene) were used to transiently achieve the knockdown of the protein. Cells were incubated for 72 h.
9
Synthesis and Characterization of PEGylated BODIPY Probe
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
γ‐Benzyl l ‐glutamate was purchased from Chuo Kaseihin (Tokyo, Japan) and MG132 (Z‐Leu‐Leu‐Leu‐al) was purchased from Sigma‐Aldrich (St. Louis, MO, USA). Bis (trichloromethyl) carbonate (triphosgene) was purchased from Tokyo Kasei Kogyo (Tokyo, Japan). Both DMF and DMAc were purchased from Wako Pure Chemicals Industries (Tokyo, Japan). α‐Methoxy‐ω‐amino‐polyethylene glycol (CH3O‐PEG‐NH2; molecular weight, 12 000 Da) was purchased from NOF (Tokyo, Japan). BODIPY TR cadaverine was purchased from Invitrogen (Carlsbad, CA, USA). Blocking One Buffer was purchased from Nacalai Tesque (Tokyo, Japan), and DMEM was purchased from Sigma‐Aldrich. Anti‐hScrib and anti‐p53 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Alexa Fluor 488‐conjugated donkey anti‐mouse IgG and Alexa Fluor 555‐conjugated goat anti‐rabbit IgG were purchased from Invitrogen.
10
Monitoring E2 and SIRT1 Stability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
N/Tert-1 cells expressing stable E2-S23A were plated on mitomycin C-treated J2 feeders in KSFM media. Next day, the cells were treated with 10 μM MG132 (Z-Leu-Leu-Leu-al; Sigma, catalog no. C2211) and harvested at 0, 2, 4, 6, and 8 h posttreatment and immunoblotting was carried out to detect E2, SIRT1 and GAPDH.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!