Dapagliflozin

Manufactured by Selleck Chemicals

Sourced in United States, Germany

Dapagliflozin is a sodium-glucose cotransporter 2 (SGLT2) inhibitor, a class of pharmaceutical compounds used to lower blood glucose levels in individuals with type 2 diabetes. It functions by reducing the reabsorption of glucose in the kidneys, leading to increased urinary glucose excretion.

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Lab products found in correlation

15 protocols using dapagliflozin

Phospho-β-catenin Signaling Pathway Modulation

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Canagliflozin, empagliflozin, and dapagliflozin were purchased from Selleck Chemicals (Houston, TX). MG-132, lithium chloride (LiCl), FTY720, WZB117, and phloretin were obtained from Sigma-Aldrich (St. Louis, MO) and Okadaic acid (OA) was from Cayman Chemical (Ann Arbor, MI). For immunoblotting, antibodies against phospho-β-catenin (Ser33/Ser37/Thr41), phospho-β-catenin (Ser45), β-catenin, Cyclin D1, Tubulin, Lamin B, and DDK-tag were purchased from Cell Signaling (Danvers, MA), and Glut1 and GAPDH antibodies were from Abcam (Cambridge, MA). PP2Ac, Glut3, and Actin antibodies were obtained from Millipore (Billerica, MA), Santa Cruz Biotechnology (San Diego, CA), and ProteinTech (Rosemont, IL), respectively. For experiments related to flow cytometry, Monoclonal CD133/2 (293C3)-APC and Mouse-IgG2b-APC antibodies were purchased from Miltenyi Biotech (Bergish Gladbach, Germany), and FITC anti-human CD326 (EpCAM) and Mouse-IgG2b-FITC antibodies were from BioLegend (San Diego, CA).

Hung M.H., Chen Y.L., Chen L.J., Chu P.Y., Hsieh F.S., Tsai M.H., Shih C.T., Chao T.I., Huang C.Y, & Chen K.F. (2019). Canagliflozin inhibits growth of hepatocellular carcinoma via blocking glucose-influx-induced β-catenin activation. Cell Death & Disease, 10(6), 420.

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Glucose Concentration Effects on RPTECs

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Primary human RPTCEs (ScienCell, Carlsbad, CA, USA) were used for our experiments. Low-glucose DMEM (5.5 mM D-glucose) (Thermo Fisher Scientific Inc., Rochford, IL, USA) or high-glucose DMEM (25 mM D-glucose) (Thermo Fisher Scientific Inc.) was used as a culture medium. The concentration of 5.5 mM D-glucose corresponded to the normal D-glucose concentration. To achieve equal osmolarity with the high-glucose culture medium (25 mM D-glucose), L-glucose (Sigma-Aldrich, St. Louis, MO, USA; Merck Millipore, Darmstadt, Germany) at a concentration of 19.5 mM was added to the cells that were cultured under a normal glucose concentration (5.5 mM D-glucose). Cells were cultured with or without 15 ng/mL dapagliflozin (Selleck Chemicals, Munich, Germany) for 24 h. RPTECs were cultured in 6-well plates (3 × 10⁵ cells) or 96-well plates (1 × 10⁴ cells) in a humidified atmosphere containing 5% CO₂ and at 37 °C. Each experiment was repeated three times.

Eleftheriadis T., Pissas G., Filippidis G., Efthymiadi M., Liakopoulos V, & Stefanidis I. (2022). Dapagliflozin Prevents High-Glucose-Induced Cellular Senescence in Renal Tubular Epithelial Cells. International Journal of Molecular Sciences, 23(24), 16107.

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Endothelial Cell Proliferation Assay

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M199 medium, streptomycin, penicillin, gelatin, heparin, trypsin, ethylenediaminetetraacetic acid (EDTA), sodium dodecyl sulfate (SDS), NaOH, elastase, cesium chloride, collagenase, phosphate-buffered saline (PBS), RNase, propidium iodide, glycerol, bromophenol blue, isolectin B4, and mercaptoethanol were from Sigma-Aldrich (St. Louis, MO). Matrigel and endothelial cell growth factor were from BD Biosciences (San Jose, CA). Antibodies against cyclin D1, cyclin E, cyclin A, p21, p27, platelet endothelial cell adhesion molecule-1 (PECAM-1), SGLT2, and β-actin were from Santa Cruz Biotechnology (Santa Cruz, CA), the antibody against SGLT1 was from GeneTex (Irvine, CA), and the antibody against phospho-retinoblastoma protein was from Cell Signaling Technologies (Beverley, MA). [³H]Thymidine (20 Ci/mmol) was from Perkin Elmer (Boston, MA). Canagliflozin, empagliflozin, and dapagliflozin were purchased from Selleck Chemicals (Houston, TX).

Behnammanesh G., Durante Z.E., Peyton K.J., Martinez-Lemus L.A., Brown S.M., Bender S.B, & Durante W. (2019). Canagliflozin Inhibits Human Endothelial Cell Proliferation and Tube Formation. Frontiers in Pharmacology, 10, 362.

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SGLT2 Inhibitors Improve T2DM in Mice

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A total of forty 5-week-old C57/B6 male mice were fed with a 60% high-fat diet for two weeks to induce insulin resistance. The T2DM mouse model was established by intraperitoneal injection of low-dose streptozotocin (STZ) (25 mg/kg) for 5 consecutive days. After 2 weeks, 24 mice with blood glucose levels higher than 16.7 mmol/L were considered to indicate type 2 diabetes mellitus (T2DM). Canagliflozin, dapagliflozin, and empagliflozin (Selleck Chemicals, Houston, TX) were mixed in a 60% high-fat diet separately and fed to T2DM mice. All mice were assigned to as following four groups: 1) T2DM without treatment (DM group); 2) T2DM treated with Canagliflozin (100 mg/kg/d) (DM+Cana group); 3) T2DM treated with dapagliflozin (10 mg/kg/d) (DM+Dapa group); and 4) T2DM treated with empagliflozin (10 mg/kg/d) (DM+Empa group). After 10 weeks, femoral tissues and plasma samples were collected. The ethics committee of Chongqing University Central Hospital has approved all animal experiments in accordance with the Declaration of Helsinki.

Song P., Chen T., Rui S., Duan X., Deng B., Armstrong D.G., Ma Y, & Deng W. (2022). Canagliflozin promotes osteoblastic MC3T3-E1 differentiation via AMPK/RUNX2 and improves bone microarchitecture in type 2 diabetic mice. Frontiers in Endocrinology, 13, 1081039.

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Protein Expression and Lipid Metabolism

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Protein expression was assessed by immunoblot analysis of cell lysates (20–60 μg) in RIPA buffer in the presence of the following antibodies: including anti-p-AMPK (Affinity, AF3423), anti-AMPK (Affinity, AF6423), anti-p-ACC1 (Cell Signaling Technology, 11818s), anti-ACC1(Cell Signaling Technology, 4190s), anti-ACOX1 (Santa Cruz Biotechnology, sc-517306), anti-p-mTOR (Affinity, AF3308), anti-mTOR (Affinity, AF6308), anti-SGLT2 (Abcam, ab37296), anti-LC3B (Cell Signaling Technology, 12741S), anti-Beclin1 (Cell Signaling Technology, 4122s), anti-p62/SQSTM1 (Proteintech, 18420-1-AP), and anti-GAPDH (Abcam, ab8245).
Unless otherwise specified, all chemicals were purchased from Sigma-Aldrich (St. Louis, MO, United States). Dulbecco's modified Eagle’s medium (DMEM), fetal bovine serum (FBS, Gibco) were obtained from Gibco; palmitate (PA) was obtained from Sigma-Aldrich (P9767); compound C (Comp C) was purchased from AbMole (M2238), chloroquine (CQ, S4157) and dapagliflozin (S1548) were bought from Selleck; and Oil Red O and triglyceride detection kit (G1262; BC0625) were obtained from Solarbio.

Li L., Li Q., Huang W., Han Y., Tan H., An M., Xiang Q., Zhou R., Yang L, & Cheng Y. (2021). Dapagliflozin Alleviates Hepatic Steatosis by Restoring Autophagy via the AMPK-mTOR Pathway. Frontiers in Pharmacology, 12, 589273.

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Glucose Transport Modulation in hCPCs

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hCPCs were maintained in Ham’s F12 medium (Hyclone, Logan, UT, USA) containing 10% fetal bovine serum (FBS, Gibco, CA, USA), 1× penicillin/streptomycin (PS, Welgene, Daegu, Republic of Korea), 2.5 U of human erythropoietin (hEPO, R&D Systems, Minneapolis, MN, USA), 10 ng/mL of basic human recombinant fibroblast growth factor (bFGF, Peprotech, Rocky Hill, NJ, USA), and 0.2 mM/L glutathione (Sigma-Aldrich, St. Louis, CA, USA). hCPCs cultured in growth medium containing d-glucose (Sigma-Aldrich) and Fasentin (Sigma-Aldrich) was used as a GLUT1 blocker (Wood et al., 2008 (link)), and dapagliflozin (Selleckchem, Houston, TX, USA) was used as an SGLT2 blocker. Both blockers were used to co-treat hCPCs cultured in medium supplemented with 25 mM d-glucose.

Choi H.Y., Park J.H., Jang W.B., Ji S.T., Jung S.Y., Kim D.Y., Kang S., Kim Y.J., Yun J., Kim J.H., Baek S.H, & Kwon S.M. (2016). High Glucose Causes Human Cardiac Progenitor Cell Dysfunction by Promoting Mitochondrial Fission: Role of a GLUT1 Blocker. Biomolecules & Therapeutics, 24(4), 363-370.

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Glucose Uptake Measurement Protocol

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Keratinocyte-SFM (serum-free medium) was obtained from Gibco (Grand Island, NY, USA). 2-NBDG was from Invitrogen (Carlsbad, CA, USA). Dapagliflozin and Phlorizin were purchased from Selleck (USA), N-Methyl-d-glucamine and all other chemicals were from Sigma-Aldrich (St. Louis, MO, USA).

Lu Y.T., Ma X.L., Xu Y.H., Hu J., Wang F., Qin W.Y, & Xiong W.Y. (2018). A Fluorescent Glucose Transport Assay for Screening SGLT2 Inhibitors in Endogenous SGLT2-Expressing HK-2 Cells. Natural Products and Bioprospecting, 9(1), 13-21.

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Cellular Metabolic Pathway Regulation

Cited 2 times

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Canagliflozin, dapagliflozin and empagliflozin were from Selleckchem;
phlorizin, phloretin, metformin, phenformin, AICAR and 2-dinitrophenol from
Sigma. A769662 was synthesized as described (23 (link)). Antibodies against phospho-Thr172 on AMPK-α (pT172,
#2531) were from Cell Signaling Technology. In Fig. 8, Antibodies against
phospho-ACC (pACC, #3661) and total ACC (#3676) were from Cell Signaling
Technology. In other Figures, total ACC was detected using streptavidin directly
conjugated to 800 nm flourophore (Rockland immunochemicals), and pACC (14 (link)) and total AMPK-α (24 (link)) antibodies were as previously
described. Anti-GLUT1 (#325510) was from Abcam and anti-SGLT2 (sc-47402) from
Santa Cruz.

Hawley S.A., Ford R.J., Smith B.K., Gowans G.J., Mancini S.J., Pitt R.D., Day E.A., Salt I.P., Steinberg G.R, & Hardie D.G. (2016). The Na+/glucose co-transporter inhibitor canagliflozin activates AMP-activated protein kinase by inhibiting mitochondrial function and increasing cellular AMP levels. Diabetes, 65(9), 2784-2794.

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Dapagliflozin Treatment in Diabetic Rats

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All animal protocols were approved by the Emory University Institutional Animal Care and Use Committee. Male Sprague-Dawley rats (Charles River Laboratories, Wilmington, MA), weighing 150–200 g, received free access to water and standard rat chow (Teklad diet # 5001) containing 23% protein. Rats were made diabetic by injection of streptozotocin (STZ; 60 mg/kg) into the tail vein. Hyperglycemia was verified 24–48 h after injection using a Lifescan Ultra II glucometer. Dapagliflozin (1 mg/kg/day; Selleck Chemicals, Houston, TX) was started 2 days after STZ injection. Dapagliflozin was dissolved in 1% hydroxypropyl methylcellulose. The dose per animal was delivered in 2 ml of this solution by gavage for 7 days or 14 days.

Chen L., LaRocque L., Efe O., Wang J., Sands J.M, & Klein J.D. (2016). Effect of Dapagliflozin Treatment on Fluid and Electrolyte Balance in Diabetic Rats. The American journal of the medical sciences, 352(5), 517-523.

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Pharmacological Inhibitors in Cancer Research

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CB-839 (S7655), BPTES (S7753), BAY-876 (S8452), AZD3965 (S7339), CPI613 (S2776), Compound 3 k (S8616), NCT503 (S8619), AG221 (S8205), NLG-8189 (S7756), IACS-10759 (S8731), Dapagliflozin (S1548), 2-DG (S4701) and DHEA (S2604) were purchased from Selleck Chemicals. ND-646 (HY-101842) was purchased from MedChemExpress. V-9302 (1855871-76-9) was purchased from Probechem Biochemicals. N-acetyl cysteine (NAC) was purchased from Sigma. Antibody against HSP90 (sc-13119) was purchased from Santa Cruz Biotechnology. Antibody against two different splice forms of GLS, KGA/GAC, (12855–1-AP) was purchased from Proteintech. Antibody against γH2AX (#9718) was purchased from Cell Signaling. Antibodies against Ki67 (ab15580) and Cleaved caspase-3 (ab2303) were from Abcam.

Jin H., Wang S., Zaal E.A., Wang C., Wu H., Bosma A., Jochems F., Isima N., Jin G., Lieftink C., Beijersbergen R., Berkers C.R., Qin W, & Bernards R. (2020). A powerful drug combination strategy targeting glutamine addiction for the treatment of human liver cancer. eLife, 9, e56749.

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