Plcγ1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

PLCγ1 is a protein that plays a key role in cellular signal transduction. It functions as a phospholipase enzyme, responsible for the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) into the secondary messengers inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG).

Automatically generated - may contain errors

Lab products found in correlation

Dnp human serum albumin, by Merck Group (3 mentions) Erk1 2, by Santa Cruz Biotechnology (2 mentions) Actin, by Merck Group (2 mentions) β catenin, by Santa Cruz Biotechnology (2 mentions) Monoclonal dinitrophenol dnp specific ige, by Merck Group (1 mentions) P38 thr180 tyr182, by Cell Signaling Technology (1 mentions) Toluidine blue o, by Merck Group (1 mentions) Image pro plus software 6, by Media Cybernetics (1 mentions) Gapdh, by Southern Biotech (1 mentions) Pi3k p85, by Santa Cruz Biotechnology (1 mentions) Histone h3, by Bioss Antibodies (1 mentions) Qproteome cell compartment kit, by Qiagen (1 mentions) β actin, by Abcepta (1 mentions) E cadherin, by Santa Cruz Biotechnology (1 mentions) Complete protease inhibitor cocktail tablet, by Roche (1 mentions) Bicinchoninic acid protein assay kit, by Dingguo (1 mentions) Rabbit polyclonal antibody, by Santa Cruz Biotechnology (1 mentions) Mouse anti dinitrophenyl dnp ige, by Merck Group (1 mentions) Antibodies for phosphotyrosine, by Merck Group (1 mentions) An antibody against β actin, by Merck Group (1 mentions)

7 protocols using plcγ1

Investigating Mast Cell Signaling Pathways

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Dasatinib (Fig. 1A) was purchased from Selleckchem (Houston, TX, USA) and 4-Amino-5-(4-chlorophenyl)-7-(dimethylethyl) pyrazolo[3,4-d] pyrimidine (PP2) was obtained from Calbiochem (La Jolla, CA, USA). Monoclonal dinitrophenol (DNP)-specific IgE, DNP-Human serum albumin (HSA), Evans blue, toluidine blue O and cetrizine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against the phosphorylated forms of Syk (Tyr32), LAT (Tyr191), PLCγ1 (Tyr783), Akt (Ser473), Erk1/2 (Thr202/Try204), JNK (Thr183/Try185) and p38 (Thr180/Tyr182) were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA). All antibodies were used diluted 1:1000, unless otherwise stated. Antibodies against total forms (dilution fold, 1:100) of Lyn (catalog no. sc-7274), Fyn (catalog no. sc-434), Syk, PLCγ1, Erk1/2, JNK and p38 were obtained from Santa Cruz Biotechnology (Dallas, TX, USA) and LAT (dilution fold, 1:1000) and Actin (dilution fold, 1:5000) antibodies were from EMD Millipore Corporation (Billerica, MA, USA). The media and reagents used for cell culture were from Welgene (Gyeongsangbuk-do, Korea) and Gibco/Life Technologies, Inc (Rockville, MD, USA).

Lee D., Park Y.H., Lee J.E., Kim H.S., Min K.Y., Jo M.G., Kim H.S., Choi W.S, & Kim Y.M. (2020). Dasatinib Inhibits Lyn and Fyn Src-Family Kinases in Mast Cells to Suppress Type I Hypersensitivity in Mice. Biomolecules & Therapeutics, 28(5), 456-464.

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Cell Lysate Extraction and Protein Analysis

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Total cell lysates were isolated by radioimmunoprecipitaion assay (RIPA) lysis buffer (50 mM HEPES, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% SDS) containing 1% PMSF and Complete Protease Inhibitor Cocktail tablets (Roche, Indianapolis, IN). Plasma membrane and nuclear lysates were extracted using the Qproteome Cell Compartment Kit (Qiagen, Leipzig, Germany). Protein concentrations were determined by a bicinchoninic acid protein assay kit (Beijing Dingguo Changsheng Biotechnology, Beijing, China). Equal amounts of protein were used for Western blot analysis with appropriate antibodies against p120, PLC-γ1, E-cadherin, β-catenin, integrin α2 (plasma membrane marker), PI3K p85, or nucleolin (Santa Cruz), PIKE or PKC-ζ (Millipore, Billerica, MA), phosphorylated nucleolin (p-nucleolin, Abcam, Cambridge, MA), histone H3 (nuclear marker, Bioss, Beijing, China), GAPDH (Southern Biotechnology Associates, Birmingham, AL), or β-actin (Abgent, San Diego, CA). Quantifications of proteins within the bands of interest were performed using Image-pro plus software 6.0 (Media Cybernetics). Each experiment was repeated 3 times.

Li L., Ji S., Shrestha C., Jiang Y., Liao L., Xu F., Liu Z., Bikle D.D, & Xie Z. (2020). p120-catenin suppresses proliferation and tumor growth of oral squamous cell carcinoma via inhibiting nuclear phospholipase C-γ1 signaling. Journal of cellular physiology, 235(12), 9399-9413.

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Antibody-Mediated Signaling Pathway Analysis

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Mouse anti-dinitrophenyl (DNP) IgE and DNP-human serum albumin (HSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies used were used: Rabbit polyclonal antibodies specific for phospho-IκB, IKKα/β, ERK1/2, JNK, PLCγ1, p38, β-actin, and total form for IκB, ERK1/2, JNK, p38, and 5-LO were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Rabbit polyclonal antibodies against NF-κB p65, Syk, LAT, PLCγ1, phospho-cPLA₂ (Ser505), IKKα/β and lamin B as well as secondary goat anti-rabbit IgG-HRP and rabbit anti-goat IgG-HRP antibodies, total Syk, total LAT, and Bay 61-3606 were purchased from Santa Cruz Biotechnology (Dallas, TX, USA) and antibodies for phosphotyrosine was purchased from Millipore (Millipore, Billerica, MA, USA). The enzyme immnoassay (EIA) kits for PGD₂ and LTC₄ were purchased from Cayman Chemicals (Ann Arbor, MI, USA).

Lu Y., Li X., Park Y.N., Kwon O., Piao D., Chang Y.C., Kim C.H., Lee E., Son J.K, & Chang H.W. (2014). Britanin Suppresses IgE/Ag-Induced Mast Cell Activation by Inhibiting the Syk Pathway. Biomolecules & Therapeutics, 22(3), 193-199.

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Culturing MKN28 Gastric Carcinoma Cells

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The MKN28 human gastric carcinoma cells obtained from the American type culture collection (Manassas, VA, USA) were cultured in RPMI medium (Invitrogen Corp, Carlsbad, CA, USA) supplemented with 10% (v/v) fetal bovine serum (FBS) (GIBCO BRL, Grand Island, NY, USA), 1 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified atmosphere of 95% air and 5% CO₂. Antibodies against TRAIL, DR4, DR5, Fas, FasL, XIAP, a cellular inhibitor of apoptosis protein-1 (cIAP-1), cIAP-2, caspase-3, -8, -9, Bcl-2, Bax, Flip_L, Flip_S, PARP, β-catenin, and PLCγ1 were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). An antibody against β-actin was from Sigma (Beverly, MA, USA). Caspase activity assay kits were obtained from R&D Systems (Minneapolis, MN, USA). An enhanced chemiluminescence (ECL) kit was purchased from Amersham (Arlington Heights, IL, USA). 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) were obtained from Calbiochem (San Diego, CA, USA). All other chemicals not specifically cited here were purchased from Sigma–Aldrich (St. Louis, MO, USA). All these solutions were stored at −20 °C. Stock solutions of 4,6-diamidino-2-phenylindole (DAPI, 100 μg/mL) and propidium iodide (PI, 1 mg/mL) were prepared in phosphate-buffered saline (PBS).

Park C., Lee W.S., Go S.I., Jeong S.H., Yoo J., Cha H.J., Lee Y.J., Kim H.S., Leem S.H., Kim H.J., Kim G.S., Hong S.C, & Choi Y.H. (2021). Apoptotic Effects of Anthocyanins from Vitis coignetiae Pulliat Are Enhanced by Augmented Enhancer of the Rudimentary Homolog (ERH) in Human Gastric Carcinoma MKN28 Cells. International Journal of Molecular Sciences, 22(6), 3030.

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CYC116 Signaling Pathway Analysis

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CYC116 [4-(2-amino-4-methyl-1,3-thiazol-5-yl)-N-[4-(morpholin-4-yl)phenyl] pyrimidin-2-amine)] (Fig. 1) was obtained from Selleckchem (catalog no. S1171; Houston, TX, USA). Monoclonal dinitrophenyl (DNP)-specific monoclonal IgE (catalog no. D8406), DNP-human serum albumin (HSA, catalog no. A6661), calcium ionomycin (catalog no. I0634), Evans blue (catalog no. E2129), cetirizine (catalog no. C3618), and toluidine blue (catalog no. 198161) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies for the detection of phosphorylation of ZAP70/Syk (catalog no. 2701), LAT (catalog no. 3584), PLCγ1 (catalog no. 2821), ERK1/2 (catalog no. 9106), JNK (catalog no. 9251), p38 (catalog no. 9211), and Akt (catalog no. 9271) were purchased from Cell Signaling Technology Inc (Danvers, MA, USA). Antibodies against Lyn (catalog no. sc-7274), Fyn (catalog no. sc-434), Syk (catalog no. sc-51703), and PLCγ1 (catalog no. sc-7290) were from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against LAT (catalog no. 06-807) and Actin (catalog no. MAB1501) were purchased from EMD Millipore (Billerica, MA, USA). Cell culture reagents, including Roswell Park Memorial Institute (RPMI) 1640 medium and fetal bovine serum (FBS) were purchased from GIBCO/Life Technologies Inc (Rockville, MD, USA).

Park Y.H., Kim H.W., Kim H.S., Nam S.T., Lee D., Lee M.B., Min K.Y., Koo J., Kim S.J., Kim Y.M., Kim H.S, & Choi W.S. (2018). An Anti-Cancer Drug Candidate CYC116 Suppresses Type I Hypersensitive Immune Responses through the Inhibition of Fyn Kinase in Mast Cells. Biomolecules & Therapeutics, 27(3), 311-317.

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PDGF and EGF Signaling Pathway

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Chemicals were purchased from Sigma-Aldrich, UK. Human PDGF was purchased from R&D Systems. Recombinant EGF was purchased from Peprotech, UK. Antibodies were purchased as follows: pAKT (Thr308), pAKT (Ser473), pPLCγ1 (Y783), total AKT and PDK1, pERK1/2 T202/Y204 and total ERK1/2 from Cell Signaling Technology, GAPDH from Abcam, PLCγ1 from Santa Cruz Biotechnology. Glass-bottomed 35-mm dishes were obtained from MatTek Corporation. InsP₅ and 2-O-Bn-InsP₅ were synthesised as previously reported21 (link)28 (link). Each compound was purified to homogeneity by ion-exchange chromatography on Q-Sepharose Fast Flow resin and used as the triethylammonium salt, which was fully characterized by ³¹P and ¹H spectroscopy and accurately quantified by total phosphate assay.

Raimondi C., Calleja V., Ferro R., Fantin A., Riley A.M., Potter B.V., Brennan C.H., Maffucci T., Larijani B, & Falasca M. (2016). A Small Molecule Inhibitor of PDK1/PLCγ1 Interaction Blocks Breast and Melanoma Cancer Cell Invasion. Scientific Reports, 6, 26142.

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Immunoblotting of Signaling Molecules

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Lipofectamine 2000 was obtained from Life Technologies (Carlsbad, CA, USA). Recombinant P3C, FSL-1, poly I:C, LPS, and IL-1β were obtained from Invivogen (San Diego, CA, USA). Antibodies against p65, c-Rel, β-actin, lamin A/C, hNRPA1, Sam68, PLCγ1, JNK1/2 and ERK1/2 were from Santa Cruz Biotechnology (San Diego, CA, USA). Antibodies detecting phosphorylated forms of ERK1/2 and JNK1/2 were from Cell Signaling Technologies (Danvers, MA, USA).

Tomalka J.A., de Jesus T.J, & Ramakrishnan P. (2016). Sam68 is a regulator of Toll-like receptor signaling. Cellular and Molecular Immunology, 14(1), 107-117.

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