Pkg 1

Manufactured by Cell Signaling Technology

Sourced in United States

PKG-1 is a laboratory product offered by Cell Signaling Technology. It functions as a recombinant protein that can be utilized in various research applications.

Automatically generated - may contain errors

Lab products found in correlation

Gapdh, by Cell Signaling Technology (2 mentions) Ab138492, by Abcam (1 mentions) Histone 3, by Merck Group (1 mentions) Active β catenin, by Merck Group (1 mentions) Ab154841, by Abcam (1 mentions) α tubulin, by Merck Group (1 mentions) P akt ser473, by Cell Signaling Technology (1 mentions) Acetyl histone h3 lys14, by Merck Group (1 mentions) P vasp ser239, by Cell Signaling Technology (1 mentions) Smad3, by Cell Signaling Technology (1 mentions) P smad3, by Cell Signaling Technology (1 mentions) α sma, by Merck Group (1 mentions) Image lab software, by Bio-Rad (1 mentions) Sequi blot pvdf membrane, by Bio-Rad (1 mentions) Subcellular protein fractionation kit for tissue, by Thermo Fisher Scientific (1 mentions) Pgc 1α, by Abcam (1 mentions) Lamin b1, by Cell Signaling Technology (1 mentions) Total oxphos, by Abcam (1 mentions) Ripa buffer, by Merck Group (1 mentions) Protease and phosphatase inhibitor, by Roche (1 mentions)

5 protocols using pkg 1

Histone Extraction and Western Blot Analysis

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Cells and tissues were lysed in RIPA buffer. Histones were extracted as described below. Samples were subjected to western blot analysis as reported [20 (link),117 (link)]. Primary antibodies were: α-tubulin (T6074, Sigma-Aldrich), SMAD3 (C67H9, Cell Signaling, Danvers, MA, USA), pSMAD3 (C25A9, Cell Signaling), α-SMA (F3777, Sigma-Aldrich), VASP (3112, Cell Signaling), pVASP (Ser239) (3114, Cell Signaling), GUCY1B3, sGC1β (ab15484) (ab154841, Abcam), PKG-1 (3248, Cell Signaling), histone-3 (05-928, Millipore), Acetyl-Histone H3 (Lys14) (06-911, Millipore), pAKT (Ser473) (9271, Cell Signaling), AKT (9272, Cell Signaling), active β-catenin (05-665, Millipore), collagen-I (ab138492, Abcam, USA), and DKK1 (AF1096, R&D Systems). For quantification of immunoblot signals we used the Image Lab Software from Bio-Rad Laboratories.

Claveria-Cabello A., Colyn L., Uriarte I., Latasa M.U., Arechederra M., Herranz J.M., Alvarez L., Urman J.M., Martinez-Chantar M.L., Banales J.M., Sangro B., Rombouts K., Oyarzabal J., Marin J.J., Berasain C., Avila M.A, & Fernandez-Barrena M.G. (2020). Dual Pharmacological Targeting of HDACs and PDE5 Inhibits Liver Disease Progression in a Mouse Model of Biliary Inflammation and Fibrosis. Cancers, 12(12), 3748.

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Cardiac Protein Profiling and Western Blotting

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Whole-cell protein extracts were prepared by homogenizing cardiac tissue in RIPA Buffer (Sigma), supplemented with protease and phosphatase inhibitors (Roche Inc. Basel, Switzerland). Nuclear extracts were obtained by using the Subcellular Protein Fractionation Kit for Tissues (Thermo Scientific). Electrophoresis was performed on 3–8% gradient Nu-Page Tris-Acetate or 10% Nu-Page Bis-Tris gels and transferred to 0.2 μm Sequi-Blot PVDF membranes (Bio-Rad). The membranes were blotted with total Creb1 (Cell Signaling, Danvers, MA, USA), PKG1 (Cell Signaling), total OXPHOS (Abcam, Cambridge, UK), PGC1α (Abcam), TBP, Lamin B1, and GAPDH (Cell Signaling) antibodies. Densitometric analysis of Western blots was performed using Adobe Photoshop (Adobe Systems, San Jose, CA, USA). For representative individual gel/blots, the reference proteins (laminB1 and TBP) are shown immediately below the proteins of interest.

Harris M.P., Zeng S., Zhu Z., Lira V.A., Yu L., Hodgson-Zingman D.M, & Zingman L.V. (2023). Myokine Musclin Is Critical for Exercise-Induced Cardiac Conditioning. International Journal of Molecular Sciences, 24(7), 6525.

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Quantitative Immunoblotting Analysis of Mitochondrial Proteins

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Immunoblotting was performed as described previously [4 (link)]. Protein concentrations in mitochondria and homogenates were determined using a Bradford assay kit (Bio-Rad, Hercules, CA, USA). Equal amounts of mitochondrial and homogenate proteins from the different experimental groups were resolved on 10% SDS-PAGE and transferred onto Amersham Hybond ECL nitrocellulose membranes (GE Healthcare Bio-Sciences Corp., Pittsburgh, PA). Membranes were blocked with 5% BSA in TBS. The antibodies used in this study were against total forms of PKCɛ (1:1000), PKCδ (1:200), AT1-R (1:200), AT2-R (1:200) (Santa Cruz Biotechnology, Santa Cruz, CA), Akt (1:1000), ERK1/2 (1:1000), JNK (1:1000), p38 (1:1000), PKG-1 (1:1000), VDAC (1:1000) (Cell Signaling Technology, Danvers, MA), and GAPDH (1:10000) (Sigma-Aldrich, MO, USA). The signals were visualized by Odyssey CLx Quantitative Fluorescent Imaging System with the secondary infrared antibodies IRDye 800CW goat anti-rabbit (1:25000) and 800CW donkey anti-goat (1:25000). Results were analyzed with Image Studio Lite Software (LI-COR Biotechnology, Lincoln, NE).

Nuñez R.E., Javadov S, & Escobales N. (2018). Critical role of angiotensin II type 2 receptors in the control of mitochondrial and cardiac function in angiotensin II-preconditioned rat hearts. Pflugers Archiv : European journal of physiology, 470(9), 1391-1403.

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Protein Expression in H295R Cells

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H295R cells (ATCC, Manassas, VA) were cultured and maintained with DMEM:F12 medium supplemented with ITS Premix (Corning, NY), 2.5% Nu-serum, 100 U/mL penicillin and streptomycin. For western blotting analysis, cells lysates were collected with NP-40 containing protease inhibitors. Primary antibodies recognizing pGC-A (R&D systems, Minneapolis, MN), NPRC (Abcam, Waltham, MA), PDE2 (FabGennix, Frisco, TX), PKG I (Cell Signaling, Danvers, MA), GAPDH (Cell Signaling) and anti-IgG secondary antibodies (Santa Cruz Biotechnology, Dallas, TX) were used.

Chen Y., Iyer S.R., Nikolaev V.O., Naro F., Pellegrini M., Cardarelli S., Ma X., Lee H.C, & Burnett JC J.r. (2022). MANP Activation Of The cGMP Inhibits Aldosterone Via PDE2 And CYP11B2 In H295R Cells And In Mice. Hypertension (Dallas, Tex. : 1979), 79(8), 1702-1712.

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Comprehensive Evaluation of HCC Cell Lines

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All the human HCC cell lines including BEL-7402 and Huh7 were obtained from Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). Cells are cultured in Dulbecco's modified Eagle medium (Sigma-Aldrich, St Louis, MO, USA) replenished with 10% fetal bovine serum (Gibco, Grand Island, NY, USA) at 37 °C in a humidified incubator containing 5% CO2. Antibodies against MMP28, Notch3, Flag-tag and GAPDH were purchased from Proteintech (Chicago, IN, USA). Antibodies against ZEB-1, ZEB-2, Snail, E-cadherin, N-cadherin, Vimentin, β-catenin, Notch4, Akt, phospho-Akt (Ser473/Thr308), PKG-I, VASP and phospho-VASP (Ser239) were purchased from Cell Signaling Technology (Beverly, MA, USA).

Zhou J., Zheng X., Feng M., Mo Z., Shan Y., Wang Y, & Jin J. (2019). Upregulated MMP28 in Hepatocellular Carcinoma Promotes Metastasis via Notch3 Signaling and Predicts Unfavorable Prognosis. International Journal of Biological Sciences, 15(4), 812-825.

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