Pde4d

Manufactured by Santa Cruz Biotechnology

Sourced in United States

PDE4D is a laboratory equipment product offered by Santa Cruz Biotechnology. It functions as a phosphodiesterase enzyme that specifically targets the PDE4D isoform. This enzyme hydrolyzes the second messenger molecule cyclic AMP (cAMP), playing a role in the regulation of cellular signaling processes.

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Lab products found in correlation

3 protocols using pde4d

Western Blot Analysis of Cell Signaling

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Cells were treated for 24 h at the indicated concentrations with the respective drugs. Total protein lysates were prepared, separated, and transferred onto a polyvinylidene difluoride membrane for Western blotting as described previously [75 (link)]. Following antibodies were used: Santa Cruz Biotechnology Inc (CA, USA): PDE4D (#sc-25100), 1:200; Rap1 (#sc-65) 1:1000; deprenylated Rap1 (C-17; Rap1A, #sc-65) 1:1000. Cell signaling Technology (MA, USA): phospho-PKA substrate (#9624), 1:1000; Creb (#9104), 1:1000; phospho-Creb (Ser133; #9198), 1:1000; Epac1 (#4155), 1:1000; Integrin α5 (#4705), 1:1000; Integrin αv (#4711), 1:1000; Integrin β1 (#9699), 1:1000; Integrin β5 (#3629), 1:1000; phospho-Erk (Thr202/Tyr204; #9101), 1:1000; Erk (#9102), 1:1000; phospho-S6 (Ser240/244; #2215), 1:1000; S6 (#2317), 1:1000; phospho-Src (Tyr416; #2101), 1:1000; Src (#2109), 1:1000; Rac (#2465), 1:1000; RhoA (67B9; #2117), 1:1000, and Vinculin (E1E9V, #13901). Sigma-Aldrich: β-actin (AC-15; #A1978), 1:1000. Secondary, horseradish peroxidase-labeled antibodies from Cell Signaling Technologies were used in working dilutions of 1:10 000.

Miklos W., Heffeter P., Pirker C., Hager S., Kowol C.R., van Schoonhoven S., Stojanovic M., Keppler B.K, & Berger W. (2016). Loss of phosphodiesterase 4D mediates acquired triapine resistance via Epac-Rap1-Integrin signaling. Oncotarget, 7(51), 84556-84574.

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Western Blotting of Liver Proteins

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Liver tissue (50 mg) was lysed using RIPA buffer containing protease and phosphatase inhibitor (Sigma-Aldrich, St. Louis, MO). For nuclear protein extraction, an EpiSeeker Nuclear Extraction Kit from Abcam (Abcam, Cambridge, MA) was used. Proteins (25 μg) were analysed by SDS-polyacrylamide gel electrophoresis using a Bio-Rad (Hercules, CA) electrophoresis system. Western blot membranes were stripped using Restore^™ PLUS Stripping Buffer (Thermo Scientific, Rockford, IL) and re-probed using another antibody. Immunoreactive bands were visualized using enhanced chemiluminescence light detection reagents (Amersham, Arlington Heights, IL). Quantification was performed with Image LabTMSoftware (BioRad, Life Science Research, Hercules, CA). PPARα, CPT1A, PGC1, SIRT1, PDE4C, PDE4D, β-actin and Histone 3 antibodies were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). PDE4B antibody was a kind gift from Dr. Marco Conti; PDE4A antibody was a gift from FabGennix Inc. (FabGennix Inc. International, Frisco, TX). The information about the source and dilution of primary antibodies is presented in Supplementary Table S1.

Avila D.V., Barker D.F., Zhang J., McClain C.J., Barve S, & Gobejishvili L. (2016). Dysregulation of hepatic cAMP levels via altered Pde4b expression plays a critical role in alcohol-induced steatosis. The Journal of pathology, 240(1), 96-107.

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Western Blot Analysis of Cardiac Proteins

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Western blot assay was performed as follows. Briefly, the protein of the H9C2 cells or cardiac tissues was extracted using a Total Protein and Neuclear-Cytosol Extraction Kit (Beyotime Institute of Biotechnology Inc., shanghai, China) following the manufacturer’s protocols. BCA assay was used to measure the protein concentration. Separated by sodium-dodecyl-sulfate polyacrylamide gel electrophoresis, and then transferred onto polyvinylidene fluoride membranes. Membranes were blocked with 5% skimmed milk, and then were incubated with primary antibodies Rabbit anti-tubulin beta antibody (Bioss, China) or β-actin (Bioss); PDE4D (Santa Cruz, USA); PDK1, p-PDK1(Ser241) Akt, p-Akt (Ser473) (CST, USA); anti-beta 1 adrenergic receptor antibody, anti-beta 2 adrenergic receptor antibody, anti-GRK2 antibody (Abcam); anti-PKA antibody (millipore); anti-CaMK2antibody(CST); β2-AR phosphorylated serine pSer346 and pSer(355,356) (sigma), overnight at 4 °C. Secondary antibody goat anti-rabbit antibody (ZSGB-BIO ORIGENE, BEIJING, China) were diluted to identify the corresponding primary antibodies. Further analysis was carried out by using an imaging densitometer (Cuene Genins) to quantify the immunoreactive bands.

Zhuang P., Huang Y., Lu Z., Yang Z., Xu L., Sun F., Zhang Y, & Duan J. (2016). cAMP-PKA-CaMKII signaling pathway is involved in aggravated cardiotoxicity during Fuzi and Beimu Combination Treatment of Experimental Pulmonary Hypertension. Scientific Reports, 6, 34903.

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