Sml0843

Manufactured by Merck Group

Sourced in United States

SML0843 is a laboratory equipment product manufactured by Merck Group. It serves as a precision instrument for various scientific applications. The core function of SML0843 is to perform accurate measurements and analysis within a controlled laboratory environment. No further details can be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

11 protocols using sml0843

Pharmacokinetic Study of ISRIB in Rats

Cited 5 times

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Trans-N,N′-(Cyclohexane-1,4-diyl)bis(2-(4-chlorophenoxy) acetamide (ISRIB, Sigma, SML0843) was dissolved in dimethyl sulfoxide (DMSO) with gentle warming and diluted in polyethylene glycol 400 (PEG400) or saline before injection; 1:1 DMSO and PEG400 or 1% v/v solution of DMSO in saline was used as vehicle control. The choice of dose and timing of ISRIB administration was based on previous reports [14 (link), 17 (link), 18 (link), 24 (link), 32 –34 (link)] and our study of the pharmacokinetics of ISRIB in live rats (see below). 4-(3-phosphonopropyl)piperazine-2-carboxylic acid ((±)-CPP, Alomone, C-175) and 3-((2-methyl-1,3-thiazol-4-yl)ethynyl)pyridine hydrochloride (MTEP hydrochloride, Abcam, ab120035) were prepared in distilled water and diluted with saline to the required concentration.

Hu Z., Yu P., Zhang Y., Yang Y., Zhu M., Qin S., Xu J.T., Duan D., Wu Y., Wang D., Rowan M.J, & Hu N.W. (2022). Inhibition of the ISR abrogates mGluR5-dependent long-term depression and spatial memory deficits in a rat model of Alzheimer’s disease. Translational Psychiatry, 12, 96.

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Regulation of Stress Granule Formation

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ISRIB (200 nM; Sigma SML0843) and cycloheximide (100 µg/ml; Sigma C4859) treatment was performed for 30 min before adding sodium arsenite (0.5 mM; Sigma 35000–1 L-R) or blue light. For sodium arsenite treatment, medium was changed to medium containing 0.25 mM or 0.5 mM sodium arsenite for 30 or 45 min as indicated in figure legends. For heat shock treatment, cells were transferred to a 42°C humidified incubator with 5% CO₂ for 1 hr.

Zhang P., Fan B., Yang P., Temirov J., Messing J., Kim H.J, & Taylor J.P. (2019). Chronic optogenetic induction of stress granules is cytotoxic and reveals the evolution of ALS-FTD pathology. eLife, 8, e39578.

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Tunicamycin-induced ER stress response

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HeLa cells (80% confluent, 7X10⁶ cells) in F12 media (30 mL, ThermoFisher), 10% FBS, and 1X antibiotic/antimycotic were treated with tunicamycin (Tm, 2.5 μg/mL in DMSO, 30 μL). To some samples were added Gb (50 μM in ethanol, 30 μL, Sigma-Aldrich, catalog number G110), Sephin1 (10 mg/mL in 0.1% DMSO, Sigma, catalog number SML 1356), and/or ISRIB (1 mg/mL in 0.1% DMSO, Sigma, catalog number SML 0843). After 6 hours incubation at 37°C, cells were harvested and lysed as described earlier. Each lysate (100 μg total protein) was treated with 1X Laemmli sample buffer and heated at 95°C for 1 min. The proteins in the lysates were separated by 10% SDS-PAGE gels, followed by visualization with Sypro®Ruby total protein stain or immunoblotting after transfer to PVDF membrane (Milipore Immobilon-P) using antibodies to COPS5 (Santa Cruz-SC-9074) or G3BP1 (Santa Cruz-365338). The gels images were obtained using a Typhoon imager (GE Healthcare Life Sciences).

Dedigama-Arachchige P.M., Acharige N.P, & Pflum M.K. (2018). Identification of PP1-Gadd34 substrates involved in the unfolded protein response using K-BIPS, a method for phosphatase substrate identification. Molecular omics, 14(2), 121-133.

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Intraperitoneally Administered ISRIB

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A single dose of ISRIB (SML0843, Sigma, St. Louis, MO, USA) at 2.5 mg kg⁻¹ (diluted in 6.25% DMSO and 6.25% PEG300) or vehicle was administered intraperitoneally 90 min prior to behavioral testing, protein analyses, or electrophysiological recordings.^{37 (link)}

Kabir Z., Che A., Fischer D., Rice R., Rizzo B., Byrne M., Glass M., De Marco Garcia N, & Rajadhyaksha A. (2017). Rescue of impaired sociability and anxiety-like behavior in adult cacna1c-deficient mice by pharmacologically targeting eIF2α. Molecular psychiatry, 22(8), 1096-1109.

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Optic Nerve Crush and Neuroprotective Interventions

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C57BL/6J mice, ZnT-3 conditional knockout mice (Gempharmatech Co., Ltd., Nanjing, China), and littermate controls were used in this study. Mice were housed in standard cages in a specific pathogen-free facility on a 12 h light/dark cycle with ad libitum access to food and water. Mice were randomly allocated for each experiment. Regarding the sex factor, which is generally irrelevant to eye morphology [38 (link)], both male and female mice were randomly used. Optic nerve surgeries were performed on 6 to 8 weeks old mice (average body weight, 20–26 g) under general anesthesia, as previously described [39 (link)]. Immediately after optic nerve crush (ONC), single- or combined-use of TPEN (100 μM; 616394, Millipore, Burlington, MA, USA), C16 (4, 20, 100 μM; I9785, Sigma-Aldrich, St. Louis, MO, USA), ISRIB (10, 100, 1000 μM; SML0843, Sigma-Aldrich), or vehicle was administered by intraocular injection (3 μL per eye).

Tang J., Liu Z., Han J., Xue J., Liu L., Lin J., Wu C., Zhang Q., Wu S., Liu C., Huang H., Fu Y., Li M., Zhuo Y, & Li Y. (2022). Increased Mobile Zinc Regulates Retinal Ganglion Cell Survival via Activating Mitochondrial OMA1 and Integrated Stress Response. Antioxidants, 11(10), 2001.

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Quantifying Cell Viability Under Stress

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U2AF1 WT, S34F and Q157R HEL cells were plated at
5×10⁴ cells/ml in 96-well plates and induced with 1
μg/ml doxycycline for 24 hours. Then, cells were spun down at
500×g for 5 min and resuspended in RPMI 1640 containing 1
μg/ml doxycycline, 500 μM sodium arsenite (Sigma-Aldrich, Cat
#S7400) and 20 nM ISRIB (Sigma-Aldrich, Cat #SML0843) or vehicle (DMSO).
After 24 hours, cell viability measurements were obtained by flow cytometry
and by colorimetric WST-1 assay (Abcam, Cat #ab65475). For flow cytometry
analysis, cells were stained with 1.5 μl 7-aminoactinomycin D (7-AAD)
solution (STEMCELL Technologies, Cat #75001) and data were acquired by LSR
Fortessa (BD Biosciences, Yale Flow Cytometry Facility). The percentage of
viable (7-AAD⁻) cells was calculated by FlowJo software
(BD Biosciences) after debris removal. For the WST-1 assay, cells were
incubated with 10 μl WST-1 reagent at 37°C for 2 hours. The
level of cell viability was assessed measuring the absorbance at 450 nm with
a plate reader.

Biancon G., Joshi P., Zimmer J.T., Hunck T., Gao Y., Lessard M.D., Courchaine E., Barentine A.E., Machyna M., Botti V., Qin A., Gbyli R., Patel A., Song Y., Kiefer L., Viero G., Neuenkirchen N., Lin H., Bewersdorf J., Simon M.D., Neugebauer K.M., Tebaldi T, & Halene S. (2022). Precision analysis of mutant U2AF1 activity reveals deployment of stress granules in myeloid malignancies. Molecular cell, 82(6), 1107-1122.e7.

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Cellular Perturbation Assays with Pharmacological Agents

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Cells were treated for the indicated times with 20 μM CCCP (carbonyl cyanide m-chlorophenyl hydrazone; Sigma-Aldrich, C2759), 10 μM OM (oligomycin A; MedChem Express, HY-16589; Sigma-Aldrich, 75351), 20 μg/mL cycloheximide (Sigma-Aldrich, C4859), 10 μM tunicamycin (MedChem Express, HY-A0098), 200 nM ISRIB (Sigma-Aldrich, SML0843).
Cells were plated 24 h before transfection with polyethylenimine (PEI 25000, Polysciences) or turbofectin (OriGene Technologies). Cells were treated, harvested, passaged or placed on selection medium 24 h after transfection.
Transfections of siRNAs (siNTC, D-001810-10-05, Horizon Discovery; siTOMM40, M-012732-00-0010, Horizon Discovery; siTIMM23, 1299001, Thermo Fisher Scientific) were performed using Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scientific) following the manufacturer’s instructions.
For induction of shRNAs from the pLKO vector, cells were treated with 500 ng/mL doxycycline hyclate (Biomol, Cay14422-1) for 5 days.
For growth assays, cells were counted and seeded at equal numbers in triplicate wells on day 0 in DMSO (control) or 200 nM ISRIB-containing medium. Cells were passaged every 2–3 days and counted using Countess® Cell Counting Chamber Slides (Thermo Fisher Scientific, C10228). Counts shown in bar graphs represent the ratio of ISRIB-treated cells versus DMSO-treated controls.

Fessler E., Krumwiede L, & Jae L.T. (2022). DELE1 tracks perturbed protein import and processing in human mitochondria. Nature Communications, 13, 1853.

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Protein Inhibition for Cellular Stress

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Carbobenzoxy-l-leucyl-l-leucyl-l-leucinal (MG132; Sigma-Aldrich 474790, Darmstadt, Germany) and integrated stress response inhibitor (ISRIB; Sigma-Aldrich SML0843) were dissolved in DMSO and cells were treated with final concentrations as described.

Lant J.T., Kiri R., Duennwald M.L, & O’Donoghue P. (2021). Formation and persistence of polyglutamine aggregates in mistranslating cells. Nucleic Acids Research, 49(20), 11883-11899.

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Screening Neuroprotective Drugs against Hypoxia and Hypoglycemia

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To induce HX condition, nf-hSCOs were cultured in a maturation medium containing 300 µM cobalt chloride (CoCl₂; Sigma). Hypoglycemic condition (HG) was induced by replacing the medium with glucose-free neurobasal medium (Thermo Fisher Scientific, Waltham, MA) mixed with 2.5 mM Glucose solution (Thermo Fisher Scientific). A combination of CoCl₂ with hypoglycemic condition (HXHG) was induced in hypoglycemic medium containing 300 µM CoCl₂. For screening of drugs, nf-hSCOs were treated with drugs 2 days prior to exposure to HXHG conditions, and subsequently exposed to HXHG condition–containing drugs (ISRIB, 10 nM; Sigma, SML0843; rapamycin, 5 μM; Sigma, R0395; metformin, 10 mM; Sigma, PHR1084; minocyclin, 2 µM; sigma, M9511). The medium was changed daily.

Shin A., Ryu J.R., Kim B.G, & Sun W. (2023). Establishment and Validation of a Model for Fetal Neural Ischemia Using Necrotic Core-Free Human Spinal Cord Organoids. Stem Cells Translational Medicine, 13(3), 268-277.

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Astrocyte Inflammatory Response Modulation

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Astrocytes cultures were incubated in serum‐free condition for 24 h and pretreated for 30 min with UB‐SCG‐51 or p‐eIF2α signal inhibitor (Sigma, SML0843), followed by recombinant T/I/C: IL‐1α (3 ng/mL, Peprotech, Thermo Fisher), TNF‐α (30 ng/mL, R&D), C1q (400 ng/mL, R&D), or PBS for 24 h.

Griñán‐Ferré C., Jarné‐Ferrer J., Bellver‐Sanchís A., Codony S., Puigoriol‐Illamola D., Sanfeliu C., Oh Y., Lee S., Vázquez S, & Pallàs M. (2023). Novel molecular mechanism driving neuroprotection after soluble epoxide hydrolase inhibition: Insights for Alzheimer's disease therapeutics. CNS Neuroscience & Therapeutics, 30(4), e14511.

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