Skf96365

Manufactured by Abcam

Sourced in United Kingdom

SKF96365 is a selective inhibitor of TRPC (Transient Receptor Potential Canonical) channels. It is a small molecule compound that blocks the activity of TRPC channels, which are involved in calcium signaling and other cellular processes.

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Lab products found in correlation

Fura 2 am, by Thermo Fisher Scientific (1 mentions) Hrp conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Ethylene glycol tetraacetic acid (egta), by Thermo Fisher Scientific (1 mentions) Anti stim1 antibody, by Abcam (1 mentions) Tgf β, by R&D Systems (1 mentions) Anti gapdh antibody, by Kangchen Biotech (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Poly l lysine (pll), by Merck Group (1 mentions) Prolong gold, by Thermo Fisher Scientific (1 mentions) Polybrene, by Merck Group (1 mentions) Puromycin, by Merck Group (1 mentions) Escin, by Santa Cruz Biotechnology (1 mentions) Ym 58483, by Abcam (1 mentions) Anti app y188, by Abcam (1 mentions) Collagen type 1, by Merck Group (1 mentions) Polyethylenimine (pei), by Polysciences (1 mentions) Pd0325901, by Axon Medchem (1 mentions) Thapsigargin tg, by Abcam (1 mentions)

5 protocols using skf96365

TGF-β Signaling and Calcium Regulation

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All cell lines used in this work were obtained from American Type Culture Collection (Manassas, VA, USA). Cells were cultured as we previously described [42 (link)].
TGF-β was purchased from R&D systems (Minneapolis, MN, USA). SKF96365 was bought from Abcam (Abcam, UK). The calcium dye Fura 2-AM and EGTA were obtained from Invitrogen (Carlsbad, CA, USA). The antibodies against ERK1/2, p-ERK1/2 were bought from Bioworld Technology (Louis Park, MN, USA) and anti-STIM1 antibody was from Abcam (Abcam, UK). The anti-GAPDH antibody was bought from KANGCHEN BIO-TECH (Shanghai, China). HRP-conjugated goat anti-rabbit antibody was purchased from Thermo Scientific (Waltham, MA, USA).

Cheng H., Wang S, & Feng R. (2016). STIM1 plays an important role in TGF-β-induced suppression of breast cancer cell proliferation. Oncotarget, 7(13), 16866-16878.

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Calcium Signaling Pathway Analysis

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ProLong Gold and Hoechst 33342 were purchased from Life Technologies. Ionomycin (free acid) was purchased from Calbiochem. CPA, ML-9, polybrene, poly-L-lysine (PLL), fetal bovine serum (FBS), puromycin, and penicillin/streptomycin were purchased from Sigma-Aldrich. Escin was purchased from Santa Cruz. YM-58483 and SKF-96365 were purchased from Abcam. The following antibodies (Abs) were used: anti-STIM1 (HPA012123), anti-SERCA2 (S1439), anti-β-actin (A5441) were from Sigma-Aldrich, anti-APP (Y188) was from Abcam, and anti-Orai1 (G2; sc-377281) was from Santa Cruz.

Gazda K., Kuznicki J, & Wegierski T. (2017). Knockdown of amyloid precursor protein increases calcium levels in the endoplasmic reticulum. Scientific Reports, 7, 14512.

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Cell Line Characterization and Reagents

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Flp-In T-REx U2OS cells were kindly provided by Dr. Gopal Sapkota (University of Dundee); HeLa and C2C12 myoblasts were from ECACC (European Collection of Cell Cultures) and distributed by Sigma-Aldrich (St. Louis, MO, USA); All cell lines were tested for contamination before carrying out the experiments shown in this report. PD0325901 was purchased from Axon Medchem (Groningen, The Netherlands); Fura-2-acetoxymethyl ester (fura-2-AM) was from Calbiochem, a brand belonging to Merck Millipore (Darmstadt, Germany); Thapsigargin (Tg) and SKF96365 were from Abcam Biochemicals (Cambridge, UK); Polyethylenimine was purchased from Polysciences, Inc (Eppelheim, Germany); Collagen, type I, was purchased from Sigma-Aldrich (St. Louis, MO, USA). GFP-Trap resin was from Chromotek GmbH (Planegg-Martinsried, Germany).

Lopez-Guerrero A.M., Tomas-Martin P., Pascual-Caro C., Macartney T., Rojas-Fernandez A., Ball G., Alessi D.R., Pozo-Guisado E, & Martin-Romero F.J. (2017). Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains. Scientific Reports, 7, 383.

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Leukemia Cell Line Culture and Signaling

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The human leukemia cell lines Nalm-6 and Reh (DSMZ®, Deutschland) were cultured in RPMI 1640 medium containing 10% fetal bovine serum, 2 mM of L-glutamine with 5000 UI/L penicillin and 50 mg/L streptomycin (Eurobio®, France), and maintained at 37°C in a 5% CO₂ humidified atmosphere. Fura-2/AM, Rhod-2/AM, Bapta-AM, U73122, SKF96365 and thapsigargin were purchased from Abcam Biochemicals, France. Poly-D-lysine, dimethylsulfoxide (DMSO), 2APB and dexamethasone were purchased from Sigma Aldrich, France. PD98059 was purchased from Euromedex, France. Antibodies to total (Erk1/2) and phosphorylated-Erk1/2 (pErk1/2) were obtained from Cell signaling Inc, France.

Abdoul-Azize S., Dubus I, & Vannier J.P. (2017). Improvement of dexamethasone sensitivity by chelation of intracellular Ca2+ in pediatric acute lymphoblastic leukemia cells through the prosurvival kinase ERK1/2 deactivation. Oncotarget, 8(16), 27339-27352.

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Preterm Birth and TRPC3 Channel Inhibition

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Mice were randomly divided into an unfertilized group (group A), an ‘infected’ [lipopolysaccharides (LPS)-treated] preterm group (group B), a 15-day gestation (preterm cesarean section group (group C), and a full-term cesarean section group (group D).
At day 15, the pregnant mice in group B received four injections of LPS in salt solution (350 µg/kg) into the celiac artery, at intervals of 3 h. In all the groups, mice were injected with a TRPC3 channel inhibitor SKF96365 (ab120280; Abcam, Cambridge, UK) or saline (1 h after the injection of group B with LPS) and the litter delivery was observed and recorded. The mice were closely observed 24 h/day to record premature deliveries (i.e., parturition earlier than at day 19). When parturition occurred in the pregnant mice (full delivery of first offspring), the uterine tissue was immediately obtained; 10% chloral hydrate (dose 0.03 ml/kg) was used for anesthetization and the mice were then sacrificed by cervical spinal dislocation. The obtained uterine and adipose tissues were irrigated with saline solution and stored at −80°C.

Chen J., Zheng D., Cui H., Liu S., Zhang L, & Liu C. (2017). Roles and mechanisms of TRPC3 and the PLCγ/PKC/CPI-17 signaling pathway in regulating parturition. Molecular Medicine Reports, 17(1), 898-910.

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