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R mg132

Manufactured by Cayman Chemical
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(R)-MG132 is a proteasome inhibitor. It is a synthetic compound used in biochemical research applications.

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Lab products found in correlation

Carfilzomib, by Selleck Chemicals (2 mentions) Rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions) Nms 873, by Merck Group (2 mentions) S trityl l cysteine, by Merck Group (2 mentions) Gentamicin, by Thermo Fisher Scientific (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Progesterone, by Merck Group (1 mentions) Estradiol, by Merck Group (1 mentions) Bay 11 7085, by Cayman Chemical (1 mentions) Prostaglandin f2α pgf2α, by Cayman Chemical (1 mentions) Trackit 100 bp dna ladder, by Thermo Fisher Scientific (1 mentions) Turbo dna free 50 reactions, by Thermo Fisher Scientific (1 mentions) Phorbol myristate acetate (pma), by Cayman Chemical (1 mentions) 8 bromoadenosine 3 5 cyclic monophosphate 8 br camp pcr primers, by Merck Group (1 mentions) Ultrapure agarose, by Thermo Fisher Scientific (1 mentions) Ultrapure glycogen, by Thermo Fisher Scientific (1 mentions) Sybr green 2x master mix, by Thermo Fisher Scientific (1 mentions) Charcoal stripped fetal bovine serum, by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)

6 protocols using r mg132

1

Comprehensive Methods for Cell Culture and Gene Expression Analysis

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Superscript III First-Strand Synthesis System, Ultrapure Glycogen, UltraPure Agarose, and Trackit 100 BP DNA ladder were purchased from Invitrogen (Carlsbad, USA). TRizol Reagent and Turbo DNA-free 50 reactions were from Ambion (Thermo Fisher). Alien QRT-PCR Inhibitor Alert 400 Reactions were purchased from Integrated Sciences Pty (Sydney, Australia). (R)-MG132, BAY-11-7085, Phorbol Myristate Acetate (PMA), and Prostaglandin F2α (PGF2α) were obtained from Cayman Chemical Company (Michigan, USA). 8-Bromoadenosine 3′,5′-cyclic monophosphate (8-Br-cAMP), PCR primers, progesterone, and estradiol were purchased from Sigma (St Louis, USA). 2 mL 2.8 mm ceramic bead kits (CK28-R) for the Precellys homogenizer (Bertin Instruments, France) were purchased from Thermo Fischer Scientific (Melbourne, Australia). L-Glutamine, Sodium Pyruvate, Gentamicin, HEPES, Dulbecco's Modified Eagle Medium (DMEM), and Charcoal Stripped Fetal Bovine Serum were obtained from Gibco (Carlsbad, USA). SYBR Green 2x Master mix was from Applied Biosystems (Carlsbad, USA). TSA was supplied by Bio-Scientific Pty. Ltd. (Sydney, Australia).
Ilicic M., Zakar T, & Paul J.W. (2017). Modulation of Progesterone Receptor Isoform Expression in Pregnant Human Myometrium. BioMed Research International, 2017, 4589214.
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2

Tau Protein Expression and Purification

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Azure C was purchased from Fluka/Sigma (St. Louis, MO), methylene blue was purchased from EMD Chemicals (Gibbstown, NJ) and olsalazine sodium was purchased from AK Scientific (Union City, CA). Expression plasmid for the human full-length four-repeat Tau in E. coli expression vector pT7C was a kind gift of Dr. Lester I. Binder (Northwestern University). Mammalian expression constructs of Tau, HA-MARK4 WT and KD are generous gifts of Dr. Bingwei Lu (Stanford University). (R)-MG132 was purchased from Cayman Chemical (Ann Arbor, MI)
Sun W., Lee S., Huang X., Liu S., Inayathullah M., Kim K.M., Tang H., Ashford J.W, & Rajadas J. (2016). Attenuation of synaptic toxicity and MARK4/PAR1-mediated Tau phosphorylation by methylene blue for Alzheimer’s disease treatment. Scientific Reports, 6, 34784.
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3

Proteasome Inhibitor (R)-MG132 Protocol

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((R)-MG132, Cayman, Cat# 13697–1). Diluted in DMSO.
Nicholatos J.W., Francisco A.B., Bender C.A., Yeh T., Lugay F.J., Salazar J.E., Glorioso C, & Libert S. (2018). Nicotine promotes neuron survival and partially protects from Parkinson’s disease by suppressing SIRT6. Acta Neuropathologica Communications, 6, 120.
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4

Synchronizing and Enriching Mitotic Cells

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HeLa cells were first synchronized in S phase by addition of thymidine (at a final concentration of 2 mM) for 24 h. S phase cells were washed with 1×PBS to remove excess thymidine and released into fresh media (DMEM/10% FBS) for 3 h. To arrest cells in prometaphase, released cells were treated with S-trityl-L-cysteine (Sigma, 164739) (at a final concentration of 5 μM) for 12–14 h. Finally, prometaphase cells were collected by vigorous pipetting, washed with 1×PBS, and used for downstream applications, including immunoprecipitation assays and/or Western blot analyses, or frozen in liquid nitrogen and stored at −80°C for later use. For cell cycle studies, prometaphase cells were released into fresh media and collected at indicated time points. For drug inhibition studies, cells were released into media containing 2 μM carfilzomib (Selleck, PR-171), 20 μM (R)-MG132 (Cayman, 13697) and/or 10 μM NMS-873 (Sigma, SML1128) for indicated times. For depletion studies, HeLa cells were transfected with 40 nM of indicated siRNAs and a 1:400 dilution of RNAiMAX transfection reagent (Thermo Fisher, 13778150) 24 h prior to synchronization.
Mitotic enrichment of HEK 293Ts and H1s was achieved by adding STLC (at a final concentration of 5 μM) to the culture media for 14–16 h.
Oh E., Mark K.G., Mocciaro A., Watson E.R., Prabu J.R., Cha D.D., Kampmann M., Gamarra N., Zhou C.Y, & Rape M. (2020). APC/C-dependent control of gene expression and cell identity. Nature, 579(7797), 136-140.
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5

Cell Culture Protocol for Hormone and Drug Treatments

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Mouse fibroblast NIH/3T3 (derived from male), human kidney cell HEK293T (derived from female), and human liver cancer cell line PLC/PRF/5 (derived from male) and SNU-387 (derived from female) were purchased from ATCC. Cell lines were authenticated by STR profiling. SNU-387 cells were cultured using ATCC-formulated RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 100 U/mL of penicillin, and 100 μg/mL of streptomycin at 37°C in 5% CO2. Other cell lines were cultured using DMEM high glucose medium (Thermo Fisher) supplemented with 10% FBS, 100 U/mL of penicillin, and 100 μg/mL of streptomycin. For hormone treatment experiments, cells were cultured and treated in phenol red-free DMEM (Thermo Fisher) supplemented with charcoal-stripped FBS (Biowest). Proteasome inhibition was induced by treatment of 10 μM of (R)-MG132 (Cayman Chemical) for 6 h. Selective HCN blocker ZD7288 was purchased from Tocris. Thapsigargin was purchased from Millipore-Sigma. Cells were treated with ZD7288 in a complete medium for 3 days.
Zhang Y., Liu X., Sun K., Luo Y., Yang J., Li A., Kiupel M., Fenske S., Biel M., Mi Q.S., Wang H, & Xiao H. (2023). Hyperpolarization-activated cyclic nucleotide-gated cation channel 3 promotes HCC development in a female-biased manner. Cell reports, 42(10), 113157.
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6

Synchronizing and Enriching Mitotic Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
HeLa cells were first synchronized in S phase by addition of thymidine (at a final concentration of 2 mM) for 24 h. S phase cells were washed with 1×PBS to remove excess thymidine and released into fresh media (DMEM/10% FBS) for 3 h. To arrest cells in prometaphase, released cells were treated with S-trityl-L-cysteine (Sigma, 164739) (at a final concentration of 5 μM) for 12–14 h. Finally, prometaphase cells were collected by vigorous pipetting, washed with 1×PBS, and used for downstream applications, including immunoprecipitation assays and/or Western blot analyses, or frozen in liquid nitrogen and stored at −80°C for later use. For cell cycle studies, prometaphase cells were released into fresh media and collected at indicated time points. For drug inhibition studies, cells were released into media containing 2 μM carfilzomib (Selleck, PR-171), 20 μM (R)-MG132 (Cayman, 13697) and/or 10 μM NMS-873 (Sigma, SML1128) for indicated times. For depletion studies, HeLa cells were transfected with 40 nM of indicated siRNAs and a 1:400 dilution of RNAiMAX transfection reagent (Thermo Fisher, 13778150) 24 h prior to synchronization.
Mitotic enrichment of HEK 293Ts and H1s was achieved by adding STLC (at a final concentration of 5 μM) to the culture media for 14–16 h.
Oh E., Mark K.G., Mocciaro A., Watson E.R., Prabu J.R., Cha D.D., Kampmann M., Gamarra N., Zhou C.Y, & Rape M. (2020). APC/C-dependent control of gene expression and cell identity. Nature, 579(7797), 136-140.
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