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3 protocols using pe anti mouse cd117 c kit

1

Chromatin Immunoprecipitation Antibody Panel

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The following antibodies were used: Suz12 (39357, Active Motif), Histone H3K27me3 (39155, Active Motif; 07-449, Millipore), Histone H3K36me3 (Ab9050, Abcam), Histone H3K27ac (ab4729, Abcam), Histone H3 (61277, Active Motif), MYC (56058, Cell Signaling), TCF4 (ab130014, Abcam), IGF2BP1 (RN001M, MBL), TCF7L2 (sc-8631, Santa Cruz), BRD4 (A301-985A1002, Bethyl), BRD3 (A302-583A, Bethyl), BRD2 (A302-368A, Bethyl), APC (MABC202, Millipore) and β‐Actin (A3854, Sigma‐Aldrich and ab49900, Abcam). Secondary antibodies were anti‐mouse (926-32210, LI-COR), anti-rabbit (926-32211, LI-COR) and anti-goat (926-32214, LI-COR). Antibodies used for FACS were: PE anti-mouse CD117/c-Kit (Biolegend, 105808), APC anti-mouse CD11b/Mac-1 (101212, Biolegend), PE-Cy7 anti-mouse Ly-6G/Gr-1 (108416, Biolegend), PE-Cy7 anti-mouse Ly-6A/E/Sca-1 (108114, Biolegend), PerCP/Cy5.5 anti-mouse CD45.2 (109828, Biolegend), PE/Cy7 anti-mouse CD117/c-kit (105814, Biolegend), APC anti-mouse CD150 (115925, Biolegend), PE-Cy7 anti-human CD11B/MAC-1 (101215, Biolegend) and APC anti-human CD36 (336207, Biolegend).
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2

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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3

Comprehensive Immune Cell Profiling

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The following anti-mouse Lineage Cocktail antibodies: FITC anti-mCD3/ FITC/ anti-mGr-1/ FITC anti-mCD11b/ FITC anti-mCD45R (B220)/ FITC antimTer-119. In addition, APC anti-mouse IL-33Rα (st2), PE/Cy7 anti-mouse CD45, PE anti-mouse CD117 (c-Kit) were used (all purchased from Biolegend). Corresponding isotype control antibodies were used as controls.
Considering the utility of a 4-color staining panel in the following experiment, prior to evaluation of ILCs in our collected samples, FM2 control and one 4-color control negative panel sample tubes were used to obtain a valid compensation matrix and determine appropriate quadrant marker placement for the panel, also PMT voltages adjusted and optimized for evaluation of target cells.
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