S2619

Manufactured by Selleck Chemicals

Sourced in United States

S2619 is a laboratory equipment product. It is a general-purpose piece of equipment designed for use in various scientific and research settings.

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Lab products found in correlation

Doxorubicin, by Merck Group (1 mentions) Nu7026, by Merck Group (1 mentions) Ly2157299, by Selleck Chemicals (1 mentions) A6185, by Merck Group (1 mentions) 5 aza dc, by Merck Group (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Mg132, by Selleck Chemicals (1 mentions) S4441, by Merck Group (1 mentions) G7021, by Merck Group (1 mentions) D5879, by Merck Group (1 mentions)

7 protocols using s2619

Cell Line Culture and Manipulation

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LNCaP, LNCaP-LN3, C4, C4–2, C4–2B, PC-3M, PC-3M-LN4, PC-3M-Pro4, and MDA-MB453 cells were grown as previously described [30 (link), 67 (link)]. The HEK293T, PC3, and C4–2 cells were cultured in DMEM supplemented with 10% fetal calf serum, 1% penicillin, and 1% streptomycin. The DNA methylase inhibitor 5-Aza-dC (10 μM) (#A3656, Sigma), the 26S proteasome inhibitors MG132 (20 μM) (#S2619, Selleckchem), or N-acetyl-L-leucyl-L-leucyl-L-nor leucinal (LLNL) (25 μM) (#A6185, Sigma), Doxorubicin (Sigma), Talazoparib (SeleckChem), NU7026 (Sigma) were used as described in the text. TGF-β receptor type I (TGF-βRI) kinase inhibitors LY2157299 and LY363947 were bought from Selleckchem.
The expression vectors encoding DACH1 [69 (link)], Ku70 (RFP-Ku70) and Ku80 (RFP-Ku80) [70 (link)], shDACH1[71 (link)] were previously described. The EGFP-DACH1 expression plasmid was made by inserting the human DACH1 cDNA into the HindIII and BamHI sites of the pEGFP-C1 vector.

Li Z., Jiao X., Robertson A.G., Sante G.D., Ashton A.W., DiRocco A., Wang M., Zhao J., Addya S., Wang C., McCue P.A., South A.P., Cordon-Cardo C., Liu R., Patel K., Hamid R., Parmar J., DuHadaway J.B., Jones S.J., Casimiro M.C., Schultz N., Kossenkov A., Phoon L.Y., Chen H., Lan L., Sun Y., Iczkowski K.A., Rui H, & Pestell R.G. (2023). The DACH1 gene is frequently deleted in prostate cancer, restrains prostatic intraepithelial neoplasia, decreases DNA damage repair, and predicts therapy responses. Research Square.

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Hypoxia-Induced Mitochondrial Regulation

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Primary PASMCs were quiesced in serum-free medium for 24 h and then exposed to normoxia (21% O₂) and hypoxia (5% O₂) in the absence or presence of mitochondria-targeted antioxidant MitoQ (0.5 μM, HY-100116A, MCE), chetomin (50 nM, GC17405, GlpBio), MG-132 (10 μM, S2619, Selleck), H₂O₂ (50, 75, and 100 mM, Beyotime), DCA (10 mM, 347795, Sigma–Aldrich), lactate (10 mM, 71718, Sigma–Aldrich), and sodium oxamate (50 μM, GC44911, GlpBio). After indicated time periods, the relative changes in mRNA, protein, metabolites, and proliferation capacity were assessed through corresponding methods. The concentrations of these compounds were chosen according to previous studies.

Chen J., Zhang M., Liu Y., Zhao S., Wang Y., Wang M., Niu W., Jin F, & Li Z. (2022). Histone lactylation driven by mROS-mediated glycolytic shift promotes hypoxic pulmonary hypertension. Journal of Molecular Cell Biology, 14(12), mjac073.

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Protein Interaction Assays and Inhibitors

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As previously mentioned co-IP and His pull-down assays were performed [32 (link)]. Cycloheximide (CHX, 20 μg/mL, MedChemExpress) and/or MG132 (20 μM, S2619, Selleck) were added to dishes at the specified concentrations for the indicated periods, accomplished by western blotting.

Ouyang S., Zeng Z., Liu Z., Zhang Z., Sun J., Wang X., Ma M., Ye X., Yu J, & Kang W. (2022). OTUB2 regulates KRT80 stability via deubiquitination and promotes tumour proliferation in gastric cancer. Cell Death Discovery, 8, 45.

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Evaluating Myc-PTX3 Protein Stability

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MSCs or 293T cells were seeded into six‐well plates at a density of 3 × 10⁵ cells/well. The cells were treated with the lysosomal inhibitor chloroquine (50 μM; 14774; CST) or the proteasomal inhibitor MG132 (10 μM; S2619; Selleck) or MG101 (10 μM; S7386; Selleck) for another 12 hours to block the autophagy‐lysosome pathway (ALP) or UPS. Protein lysates were subsequently harvested, and the level of the Myc‐PTX3 protein was evaluated using Western blotting.

Ma M., Yang W., Cai Z., Wang P., Li H., Mi R., Jiang Y., Xie Z., Sui P., Wu Y, & Shen H. (2021). SMAD‐specific E3 ubiquitin ligase 2 promotes angiogenesis by facilitating PTX3 degradation in MSCs from patients with ankylosing spondylitis. Stem Cells (Dayton, Ohio), 39(5), 581-599.

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Optimizing Plant Seedling Transfer and Treatment Protocols

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Generally, to avoid manipulation-related effect during plant transfer from solid to liquid medium, we transferred the seedlings into liquid 1/2 MS medium and preincubated them for 12 hours before treatment.
The procedure for CRD treatment was as described [68 (link)]. The 7-DAG seedlings were collected and placed in incubation buffer (1 mM pipes [pH 6.25], 1 mM sodium citrate, 1 mM KCl, 15 mM sucrose), 200 μM CRD (C3394, Sigma-Aldrich), with or without 1 µM RALF1. The samples were infiltrated under vacuum for 3 minutes in incubation buffer, followed by incubation in 23°C for indicated times. Then, plant samples were harvested for RNA extraction.
Treatment of plants with MG132 and CHX was conducted as described [69 (link)]. Seven-day-old Col-0 and fer-4 seedlings grown on 1/2 MS agar plates were collected and transferred to liquid MS medium (with or without 1 µM RALF1) in the presence or absence of 50 μM MG132 (S2619, Selleckchem) or 100 μM CHX (01810, Sigma-Aldrich) for 2 hours. Whole-plant samples were harvested and frozen in liquid nitrogen for protein extraction and immunoblot assay.

Li C., Liu X., Qiang X., Li X., Li X., Zhu S., Wang L., Wang Y., Liao H., Luan S, & Yu F. (2018). EBP1 nuclear accumulation negatively feeds back on FERONIA-mediated RALF1 signaling. PLoS Biology, 16(10), e2006340.

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BMP2/4 and LSD1 Inhibition Effects

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Cells were pre‐treated with either active BMP2 recombinant protein (10 ng/ml, 355‐BM‐010, Bio‐Techne Ltd), active BMP4 recombinant protein (10 ng/ml, 120‐05‐ET, PeproTech), ORY‐1001 (1 μM), GSK‐LSD1 (1 μM), or DMSO for 24 h. Prior to protein extraction, each condition was split and was either co‐treated with the proteasome inhibitor MG‐132 (1 μM, S2619, Selleck Chemicals) or DMSO for 2 h. Cells were pelleted, washed with PBS, and lysed with RIPA buffer as described above. Immunoblotting was conducted according to standard procedures.

Leiendecker L., Jung P.S., Krecioch I., Neumann T., Schleiffer A., Mechtler K., Wiesner T, & Obenauf A.C. (2020). LSD1 inhibition induces differentiation and cell death in Merkel cell carcinoma. EMBO Molecular Medicine, 12(11), e12525.

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Mesangial Cell Culture and Treatments

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The mouse glomerular mesangial SV40-MES-13 cell line (Fuxiang Biotechnology Company, Shanghai, China, ATCC number CRL-1927) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 100 units/mL penicillin, 100 mg/mL streptomycin, and 10% fetal bovine serum. Confluent cells were cultured in serum-free DMEM media for 24 h before the experiments. The cells were divided into the normal group (NG, 5.56 mM), the vehicle control group (DMSO, 0.05%, D5879, Sigma, St. Louis, MO, USA), the HG group (HG, 30 mM, G7021, Sigma), the HG + low-dose AB38b group (AB38bL + HG, 2.5 μM), the HG + medium-dose AB38b group (AB38bM + HG, 5 μM), the high glucose + high-dose AB38b group (AB38bH + HG, 10 μM), the high glucose + resveratrol (5010, Sigma) positive control group (Res + HG, 10 μM), the sulforaphane positive control group (S4441, Sigma) and the MG132 positive control group (S2619, Selleckchem, Houston, TX, USA).

Du L., Wang L., Wang B., Wang J., Hao M., Chen Y.B., Li X.Z., Li Y., Jiang Y.F., Li C.C., Yang H., Gu X.K., Yin X.X, & Lu Q. (2019). A novel compound AB38b attenuates oxidative stress and ECM protein accumulation in kidneys of diabetic mice through modulation of Keap1/Nrf2 signaling. Acta Pharmacologica Sinica, 41(3), 358-372.

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