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Phosphor plcγ1

Manufactured by Cell Signaling Technology

Phosphor-PLCγ1 is a laboratory reagent that detects the phosphorylation state of the phospholipase C gamma 1 (PLCγ1) protein. PLCγ1 plays a critical role in cellular signaling pathways. The Phosphor-PLCγ1 product allows researchers to analyze the activation and regulation of PLCγ1 in various experimental systems.

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3 protocols using phosphor plcγ1

1

Dissecting PDGFR-mediated Signaling Pathways

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VSMCs were lysed in RIPA buffer and protein contents were measured using the BCA assay. Total lysates were immunoprecipitated with antibody against PDGFRβ (Santa Cruz Biotechnology, sc-432). Immunoprecipitates or cell lysates were subjected to Western blotting with antibodies to phospho-STAT1 (Invitrogen 333400), phospho-STAT3 (Cell Signaling, 9145), phospho-STAT5 (Cell Signaling, 9359), β-Actin (Cell Signaling, 4970L), phospho-Tyrosine (Millipore, 05–321), total STAT1 (Millipore, 06–501), total PLCγ1 (Cell Signaling 2822), and phosphor-PLCγ1 (Cell Signaling 14008). Primary antibodies were used at 1:1000. HRP-conjugated secondary antibodies (Jackson ImmunoResearch) were used at 1:5000.
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2

Dissecting PDGFR-mediated Signaling Pathways

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VSMCs were lysed in RIPA buffer and protein contents were measured using the BCA assay. Total lysates were immunoprecipitated with antibody against PDGFRβ (Santa Cruz Biotechnology, sc-432). Immunoprecipitates or cell lysates were subjected to Western blotting with antibodies to phospho-STAT1 (Invitrogen 333400), phospho-STAT3 (Cell Signaling, 9145), phospho-STAT5 (Cell Signaling, 9359), β-Actin (Cell Signaling, 4970L), phospho-Tyrosine (Millipore, 05–321), total STAT1 (Millipore, 06–501), total PLCγ1 (Cell Signaling 2822), and phosphor-PLCγ1 (Cell Signaling 14008). Primary antibodies were used at 1:1000. HRP-conjugated secondary antibodies (Jackson ImmunoResearch) were used at 1:5000.
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3

Characterization of Nanoparticles in Mast Cell Activation

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Aluminum oxide (Al2O3, 20 nm and 26.93 μm), titanium oxide (TiO2, 20 nm and 1 μm), iron oxide (Fe2O3, 50 nm and 1 μm) and silicon dioxide (SiO2, 20 nm and 2.34 μm) were purchased from Shanghai YunFu Nanotechnology Co., Ltd. (Shanghai, China), Fuchen Chemical Reagent Co., Ltd. (Tianjin, China), and Aladdin (Shanghai, China). Monoclonal anti-dinitrophenol (DNP)-IgE antibody, DNP-human serum albumin (HSA), and 4-nitrophenyl N-acetyl-β-D-glucosaminide were obtained from Sigma–Aldrich (St. Louis, USA). Antibodies as against phosphor-JNK, JNK, phosphor-LYN, LYN, phosphor-Plcγ1, Plcγ1, phosphor-P38, P38, phosphor-ERK1/2, ERK1/2, NF-κB, and β-actin were obtained from Cell Signaling Technology Inc. (Danvers, MA). ERK inhibitor U0126-EtOH and p38 inhibitor SB203580 were purchased from MedChem Express (Monmouth Junction, NJ, USA), PE-anti-mouse CD117 (c-kit), APC-anti-mouse FcϵRIα, and FITC-anti-mouse IgE antibodies were purchased from Biolegend (San Diego, CA, USA). Recombinant interleukin (IL)-3 and stem cell factor (SCF) were purchased from PeproTech (Cranbury, NJ, USA). Evans blue and formamide were obtained from Dalian Meilun Biotechnology Co. Ltd. (Dalian, China). The ovalbumin (OVA; grade V) was purchased from Sigma Company (St. Louis, USA), and aluminum hydroxide was purchased from Thermo Fisher Scientific (Massachusetts, USA).
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