Kx2 391

Manufactured by Selleck Chemicals

Sourced in United States

The KX2-391 is a laboratory instrument used for the analysis and measurement of various chemical and biological samples. It is designed to provide accurate and reliable data for researchers and scientists working in a wide range of fields.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Propidium iodide, by Thermo Fisher Scientific (3 mentions) Saracatinib, by Selleck Chemicals (2 mentions) Ruxolitinib, by LC Laboratories (2 mentions) Dasatinib, by Selleck Chemicals (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Recombinant human tnf α, by Thermo Fisher Scientific (1 mentions) Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions) Recombinant human pdgf bb, by Thermo Fisher Scientific (1 mentions) Bosutinib, by Selleck Chemicals (1 mentions) Peg300, by Merck Group (1 mentions) Calf intestinal alkaline phosphatase, by New England Biolabs (1 mentions) Shp099, by Novartis (1 mentions) Pdgf bb, by R&D Systems (1 mentions) Epidermal growth factor (egf), by R&D Systems (1 mentions) Tofacitinib, by Selleck Chemicals (1 mentions) Selumetinib, by Selleck Chemicals (1 mentions) Trametinib, by Selleck Chemicals (1 mentions) Pd98059, by LC Laboratories (1 mentions)

15 protocols using kx2 391

Inducing EMT through TGFβ and TNFα

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To induce EMT with TGFβ and TNFα, cells were treated with 5 ng/mL of recombinant human TGF-β1 (PeproTech) and 5 ng/mL of recombinant human TNFα (PeproTech) for the indicated time period up to 10 days. During the course of these treatments the cells were split every other day. To artificially arrest cells in S-phase MCF-10A, HCC1806 and A549 cells were treated with 1 mM thymidine for 24 hours or 7 days. Cells were seeded and treated the next day; during the 7 day treatment they were split every other day. To inhibit Sp1, MCF-10A cells were plated and treated the following day with the vehicle (0.1% DMSO) or 5 µM mithramycin A (Sp1 inhibitor –Enzo Life Sciences) for 24 hours. To inhibit SRC during EMT induction, MCF-10A cells were plated and pretreated the following day with the vehicle (0.1% DMSO) or 25 nM KX2–391 (SRC inhibitor – Selleckchem) for 1 hour before adding TGFβ and TNFα for an additional 24 hours.

Gomes A.P., Ilter D., Low V., Rosenzweig A., Shen Z.J., Schild T., Rivas M.A., Er E.E., McNally D.R., Mutvei A.P., Han J., Ou Y.H., Cavaliere P., Mullarky E., Nagiec M., Shin S., Yoon S.O., Dephoure N., Massagué J., Melnick A.M., Cantley L.C., Tyler J.K, & Blenis J. (2019). Dynamic incorporation of histone H3 variants into chromatin is essential for acquisition of aggressive traits and metastatic colonization. Cancer cell, 36(4), 402-417.e13.

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Establishing and Characterizing NPC Cell Lines

Cited 1 time

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HONE1, HK1, CNE1, CNE2, SUNE1, C666-1, and NPC43 NPC cell lines, and immortalized nasopharyngeal epithelial cell lines NP69 and NP460 were cultured as previously described [4 (link),18 (link)]. Cell lines were authenticated by and obtained from the Hong Kong NPC AoE Cell Line Repository. HK11.19 is a HONE1/chromosome 11 microcell hybrid cell line (MCH) that was established by the transfer of an additional intact human chromosome 11 into HONE1 and expressed physiological levels of THY1 (in the recipient HONE1 cells, THY1 expression is downregulated by promoter hymermethylation, and human THY1 maps to chromosome 11q22.3) [4 (link),19 (link)]. Recombinant human PDGF-BB was obtained from PeproTech (PeproTech, Rocky Hill, NJ, USA, #100-14B-10). KX2-391 (Selleckchem, Houston, TX, USA, #S2700), bosutinib (Selleckchem, Houston, TX, USA, #S1014), and saracatinib (Selleckchem, Houston, TX, USA, #1006) were prepared in DMSO at a concentration of 10 mM for an in vitro assay. For the in vivo assay, KX2-391 was first dissolved in DMSO to a concentration of 85 mg/mL, then formulated with PEG-300 (Sigma-Aldrich, St. Louis, MI, USA, #202371) and PBS before injection following the manufacturer’s instructions (the injection solution contains 4% DMSO, 30% PEG-300, and 66% PBS).

Chen L., Chau W.Y., Yuen H.T., Liu X.H., Qi R.Z., Lung M.L, & Lung H.L. (2023). THY1 (CD90) Maintains the Adherens Junctions in Nasopharyngeal Carcinoma via Inhibition of SRC Activation. Cancers, 15(7), 2189.

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Compound 11a-1: Structure-Guided Synthesis

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Compound 11a-1, 6-Hydroxy-3-iodo-1-methyl-2-(3-(2-oxo-2-((4-(thiophen-3-yl)-phenyl)amino)acetamido)phenyl)-1H-indole-5-carboxylic acid was developed and synthesized using a structure-guided and fragment-based library approach^{14 (link)}. SHP099 was obtained from Novartis. KX2-391 was obtained from Selleck Chemicals. Calf intestinal alkaline phosphatase was obtained from New England Biolabs. EGF and PDGF-BB were obtained from R&D Systems.

Kano Y., Gebregiworgis T., Marshall C.B., Radulovich N., Poon B.P., St-Germain J., Cook J.D., Valencia-Sama I., Grant B.M., Herrera S.G., Miao J., Raught B., Irwin M.S., Lee J.E., Yeh J.J., Zhang Z.Y., Tsao M.S., Ikura M, & Ohh M. (2019). Tyrosyl phosphorylation of KRAS stalls GTPase cycle via alteration of switch I and II conformation. Nature Communications, 10, 224.

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Inhibitor Compound Preparation and Storage

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Selumetinib (AZD6244, MEK inhibitor), trametinib (GSK1120212, MEK inhibitor), KX2-391 (Src inhibitor) and tofacitinib (CP-690550, JAK3 inhibitor) were purchased from Selleck Chemicals (Houston, TX, USA). PD98059 (MEK inhibitor) and ruxolitinib (INCB081424, JAK1/2 inhibitor) were purchased from LC Laboratories (Woburn, MA, USA). LY5 (STAT3 inhibitor [67 (link)]) was synthesized by Chenglong Li's Lab (College of Pharmacy, The Ohio State University). The drug compounds were dissolved in sterile dimethyl sulfoxide (DMSO) to make a 20 mM stock solution stored at −20°C.

Zhao C., Xiao H., Wu X., Li C., Liang G., Yang S, & Lin J. (2015). Rational combination of MEK inhibitor and the STAT3 pathway modulator for the therapy in K-Ras mutated pancreatic and colon cancer cells. Oncotarget, 6(16), 14472-14487.

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Kinase Inhibitor Library Screening

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A kinase inhibitor library, collection of 193 kinase inhibitors, and identified specific kinase inhibitors (HMN-214, AT7519, SNS-032, and KX2-391) were purchased from Selleckchem (Houston, TX, USA). All chemicals were dissolved in dimethyl sulfoxide (DMSO) before treatment, and the cells were treated with various concentrations. To screen small-molecule compounds that modulate luc-21-miRDREL A549 cells (1 × 10⁴ cells/well) in a 96-well plate were treated with 0.1, 0.2, 1, and 4 μM of kinase inhibitor library for 48 h. Cells were assayed with a Dual-Luciferase Reporter Assay System (Promega).

Seo Y., Kim S.S., Kim N., Cho S., Park J.B, & Kim J.H. (2020). Development of a miRNA-controlled dual-sensing system and its application for targeting miR-21 signaling in tumorigenesis. Experimental & Molecular Medicine, 52(12), 1989-2004.

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Comparative Kinase Inhibitor Evaluation

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Bosutinib (SKI-606), Dasatinib (BMS-354825), KX2-391, and Saracatinib (AZD0530) were obtained from Selleck Chemicals. PP1, PP2, Src Inhibitor-1, and SU6656 were from Sigma.

Kiris E., Burnett J.C., Nuss J.E., Wanner L.M., Peyser B.D., Du H.T., Gomba G.Y., Kota K.P., Panchal R.G., Gussio R., Kane C.D., Tessarollo L, & Bavari S. (2015). Src Family Kinase Inhibitors Antagonize the Toxicity of Multiple Serotypes of Botulinum Neurotoxin in Human Embryonic Stem Cell-Derived Motor Neurons. Neurotoxicity research, 27(4), 384-398.

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Mitochondrial Function and Cytokine Profiling

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Oligomycin, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone, antimycin, and CHX were purchased from Sigma-Aldrich (St. Louis, MO). Decitabine, monensin, MG132, KX2-391, and IACS-010759 were from Selleck (Houston, TX). Recombinant human CYTL1 was purchased from OriGene Technologies (Rockville, MD). Anti-CYTL1 antibody was purchased from Abcam (Cambridge, UK). Anti-NDUFV1, anti-BRCA1, anti-Flag, anti-GAPDH, and anti-Src antibodies were from Proteintech Group (Chicago, IL). Anti-HA, anti-CD31, anti-MDM2, anti-LDHA, anti-p-LDHA (Y10), and anti-COX4 antibodies were from Cell Signaling Technology (Beverly, MA). Anti-Tubulin, anti-β-Actin, anti-myc, and p-Src (Santa Cruz) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). Lipofectamine 3000 and Lipofectamine RNAi MAX were purchased from Thermo Fisher Scientific (Waltham, MA). MitoCheck Complex I activity assay kit (No.700930) was purchased from Cayman Chemical (Ann Arbor, Michigan). Human CYTL1 ELISA Kit was purchased from CUSABIO Life Sciences (College Park, MD).

Xue W., Li X., Li W., Wang Y., Jiang C., Zhou L., Gao J., Yu Y., Shen Y, & Xu Q. (2022). Intracellular CYTL1, a novel tumor suppressor, stabilizes NDUFV1 to inhibit metabolic reprogramming in breast cancer. Signal Transduction and Targeted Therapy, 7, 35.

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Immunofluorescence Analysis of PNMA5 and GSK-3β

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Cells pretreated with the Src protein inhibitor KX2-391 (20 nM) (Selleck, Houston, Texas, USA) or transfected with PNMA5 siRNA were fixed with paraformaldehyde for 10 minutes at room temperature, and 500 µL of blocking solution [diluted by PBS/PBST, with 10% goat serum (ZSBIO, Beijing, China) and 5% BSA] was added for one hour at 37 °C. Then, 200 µL of primary antibody (1:1,000 dilution ratio of p-PNMA5 or p-GSK-3β) was added, and the mixture was incubated overnight at 4 °C. Two hundred µL of fluorescent secondary antibody (Cy2-conjugated donkey anti-rabbit IgG, Jackson ImmunoResearch Laboratory, USA) was added for one hour at 37 °C, after which 100 µL of DAPI was added for nuclear staining at room temperature for 10 minutes. Images were acquired using a laser confocal microscope (Eclipse Ti, Nikon, Tokyo, Japan).

Qiao H., Liu J., Yang Z., Gao L., Zhang X., Lu G., Liu Y., Liu M., Ying X, & Zhang D. (2020). PNMA5 knockdown suppresses the proliferation, invasion, and metastasis of epithelial ovarian cancer cells. Translational Cancer Research, 9(5), 3691-3702.

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Cultivation and treatment of human medulloblastoma cell lines

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Human medulloblastoma cell lines (UW426, UW288 and DAOY) were provided by Dr. Corey Raffel (The Research Institute at Nationwide Children’s Hospital, USA). Normal Human Astrocytes (NHA) were purchased from ATCC (the American Type Culture Collection, Manassas, VA, USA). Cells were maintained in Dulbecco’s Modification of Eagle’s Medium (DMEM, Hyclone, USA) with 4.5 g/L, L-glutamine and sodium pyruvate (Mediatech, USA) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, USA), and 1% Penicillin/Streptomycin (P/S) (Sigma, USA) in incubators set at 37 °C and aired with 5% CO₂.
All the reagents in the study were purchased as follows: dimethyl sulfoxide (Sigma, USA), ruxolitinib (LC Laboratories, Woburn, MA, USA), tofacitinib (LC Laboratories, Woburn, MA, USA), KX2–391 (Selleckchem, Woburn, MA, USA), dasatinib (LC Laboratories, Woburn, MA, USA) and cisplatin (Sigma, USA). All the drugs were stored at −20 °C with 20 mM stock solution.

Wei J., Ma L., Li C., Pierson C.R., Finlay J.L, & Lin J. (2019). Targeting Upstream Kinases of STAT3 in Human Medulloblastoma Cells. Current cancer drug targets, 19(7), 571-582.

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Investigating Akt Signaling Pathways

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Western blots were performed as described (Yang et al., 2007 (link)) with the following antibodies: p-Akt (S473), p-Akt (T308), Akt, p-p70S6K (T389), p70S6K, RSK1, p-RSK2 (S227), RSK2, p-PDK1 (S241), PDK1, PI3K p110α, and GAPDH (Cell Signaling Technology Inc., MA, USA); Na⁺/K⁺-ATPase α1 (Abcam Inc. Cambridge, UK); and an antibody against TGEV N protein, as described (Yang et al., 2007 (link)). Ouabain (O3125, ≧95%, HPLC) was purchased from Sigma-Aldrich (St. Louis, MO, USA). p-RSK1 (T359/S363) antibody, BX795, rottlerin, GF109203X, rapamycin, and triciribine were purchased from Merck Millipore Calbiochem (Billerica, MA). LY294002, PP2, KX2-391 and SU6656 were from Selleckchem (Houston, TX, USA). Farnesyl thiosalicylic acid (FTA, a Rac1 inhibitor) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Wortmannin was purchased from Life Technologies (San Diego, CA, USA).

Yang C.W., Chang H.Y., Lee Y.Z., Hsu H.Y, & Lee S.J. (2018). The cardenolide ouabain suppresses coronaviral replication via augmenting a Na+/K+-ATPase-dependent PI3K_PDK1 axis signaling. Toxicology and Applied Pharmacology, 356, 90-97.

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