Epac2

Manufactured by Abcam

Sourced in China, United Kingdom

Epac2 is a laboratory reagent protein that functions as a guanine nucleotide exchange factor (GEF) for the Rap1 and Rap2 small GTPases. It is involved in the regulation of diverse cellular processes, including cell adhesion, cell-cell junction formation, and signal transduction.

Automatically generated - may contain errors

Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Ecl plus reagent, by Solarbio (1 mentions) Mem per plus membrane protein extraction kit, by Thermo Fisher Scientific (1 mentions) Gapdh, by ZSGB-BIO (1 mentions) Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions) β actin, by Abcam (1 mentions) Transwell chamber, by Corning (1 mentions) Trypsin, by Merck Group (1 mentions) Matrigel basement membrane matrix, by BD (1 mentions) Brain derived neurotrophic factor (bdnf), by Abcam (1 mentions) Odysseyxl system, by LI COR (1 mentions) Epac1, by Abcam (1 mentions) Trans blot turbo rapid transfer system, by Bio-Rad (1 mentions) β actin, by Merck Group (1 mentions) Occludin, by Abcam (1 mentions)

4 protocols using epac2

Membrane Protein Extraction and Western Blot

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The membrane protein was prepared using Mem-PER Plus Membrane Protein Extraction Kit (Thermo Scientific, Shanghai, China). Equal amounts of protein were separated by SDS-PAGE and transferred to NC membranes. The membranes were incubated overnight at 4 °C with antibodies against mouse GLP-1R (1:2000, Proteintech, Wuhan, China), Epac2 (1:2000, Abcam, Shanghai, China) and GAPDH (1:10,000, ZSGB Bio, Beijing, China), followed by incubation with HRP-conjugated secondary antibodies (1:10,000, Abbkine Biotech, Wuhan, China). Protein bands were visualized by ECL Plus Reagent (Solarbio, Beijing, China) and imaged on MicroChemi chemiluminescence detection system (DNR Bio Imaging System, Neve Yamin, Israel). The protein bands were analyzed using ImageJ software (NIH, Bethesda, MD, USA).

Wang Q., Zhao C., Jin L., Zhang H., Miao Q., Liu H, & Zhang D. (2018). AWRK6, a Novel GLP-1 Receptor Agonist, Attenuates Diabetes by Stimulating Insulin Secretion. International Journal of Molecular Sciences, 19(10), 3053.

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EPAC2 Overexpression and Cell Migration

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Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA), fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) and trypsin (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) were obtained. DMEM was supplemented with 10% FBS, 100 U/ml penicillin and 100 µg/ml streptomycin. Matrigel basement membrane matrix (BD Biosciences, San Jose, CA, USA) and Transwell chambers (Corning Inc., Corning, NY, USA) were obtained. EPAC2, MMP-2 and β-actin antibodies were obtained from Abcam (Cambridge, UK). EPAC2 overexpression plasmid was purchased from GenScript (Nanjing, China). Lipofectamine^® 3000 transfection reagent was obtained from Invitrogen; Thermo Fisher Scientific, Inc.

Jiang M., Zhuang Y., Zu W.C., Jiao L., Richard S.A, & Zhang S. (2019). Overexpression of EPAC2 reduces the invasion of glioma cells via MMP-2. Oncology Letters, 17(6), 5080-5086.

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Protein Quantification and Characterization

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Cells were harvested using standard techniques (cell lysis buffer containing protease inhibitors). Protein was separated by SDS PAGE using a 4–15% Tris-HCl gel and transferred to nitrocellulose membranes using a Bio-Rad Trans-Blot Turbo rapid transfer system (Bio-Rad, Hercules, CA). Primary antibodies used include BDNF, Epac1, Epac2 (Abcam, Cambridge, MA), and β-actin (Sigma-Aldrich, St. Louis, MO). Membranes were imaged on a Li-Cor OdysseyXL system and densitometry was quantified with Image Studio software. Protein blots were normalized to β-actin.

Thompson M.A., Britt RD J.r., Kuipers I., Stewart A., Thu J., Pandya H.C., McFarlane P., Pabelick C.M., Martin R.J, & Prakash Y.S. (2015). cAMP-Mediated Secretion of Brain-Derived Neurotrophic Factor in Developing Airway Smooth Muscle. Biochimica et biophysica acta, 1853(10 0 0), 2506-2514.

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Colonic Tight Junction Proteins

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Immunofluorescence was executed on colonic tissue using antibodies against ZO-1 and occludin (1:100; Abcam) to determine the changes in TJ protein levels of bowel.
Immunohistochemistry was executed on human colonic tissue using antibodies against Epac-2 and Rap-1 (1:100; Abcam).

Song X., Zuo L., Wang L., Zhu Z., Tao J., Jiang Y., Wu X., Wang Z., Nian J., Xiang P., Zhang Q., Zhao H., Liang Y., Li J., & Hu J. (2020). Rottlerin ameliorates DSS-induced colitis by improving intestinal barrier function via activation of the Epac-2/Rap-1 signaling pathway.

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