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2 protocols using goat anti vegfr1

1

VEGFR2 and NFAT Signaling in Endothelial Cells

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Antibodies: goat-anti-VEGFR2 (R&D Systems, Minneapolis, MN, USA), goat-anti-VEGFR1 (R&D Systems), rabbit-anti-ERK1/2, mouse-anti-phospho-ERK1/2 (Thr202/Tyr204), rabbit-anti-p38, rabbit-anti-phospho-p38 (Thr180/Tyr182), rabbit-anti-phospho-VEGFR2 (Tyr1175), rabbit-anti-eNOS, rabbit-anti-phospho-eNOS(Ser1177), rabbit-anti-phospho-PLCγ1(Tyr783), rabbit-anti-NFATc2 (Cell Signaling Technology, Danvers, MA, USA), mouse anti-PECAM-1 (CD31), rabbit-anti-PLCγ1 (Santa Cruz Biotechnology, Dallas, TX, USA) and mouse-anti-α-tubulin (Sigma-Aldrich, Poole, UK). Reagents: Endothelial cell growth medium (ECGM) was from PromoCell (Heidelberg, Germany). Scrambled and NFATc2 siRNA duplexes were purchased as siGENOME SMARTpools from Dharmacon (Thermo Scientific, Lafayette, USA). Recombinant human VEGF-A165 was from Genentech Inc. (San Francisco, CA, USA) and VEGF-A121 was from Promocell.
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2

VEGFR Signaling Pathway Analysis

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Antibodies: goat-anti-VEGFR1 (#AF321), goat-anti-VEGFR2 (#AF357), rabbit-anti-phospho-VEGFR2-Y1214 (#AF1766) (R&D Systems, Minneapolis, MN, USA), rabbit-anti-Akt (#9272S), rabbit-anti-phospho-Akt (S473; #4060B), rabbit-anti-ERK1/2 (#9102S), mouse-anti-phospho-ERK1/2 (T202/Y204; #9106S), rabbit-anti-neuropilin 1 (NRP1; #3725S), rabbit-anti-phospho-VEGFR2-Y951 (#4991S), rabbit-anti-phospho-VEGFR2-Y1059 (#3817S), rabbit-anti-phospho-VEGFR2-Y1175 (#2478S; Cell Signaling Technology, Danvers, MA, USA), rabbit-anti-phospho-VEGFR2-Y1054 (Clone D1W; #04-894; Merck Millipore, Watford, UK), mouse-anti-α-tubulin (Clone DM1A; #T6199; Sigma Aldrich, Poole, UK), mouse-anti-PECAM-1 (CD31; #sc-65260; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-ubiquitin (FK2; #14220; Caymen Chemical, MI, USA), mouse-anti-clathrin heavy chain antibody (X22; #ab2731; Abcam, Cambridge, UK), sheep-anti-TGN46 (#AHP500GT; AbD Serotec, Oxford, UK). Endothelial cell growth medium (ECGM) was from PromoCell (Heidelberg, Germany). Recombinant human VEGF-A165 was from Genentech Inc. (San Francisco, CA, USA), both VEGF-A121 and VEGF-A145 was from Promocell.
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