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2 protocols using anti lyve

1

Immunofluorescence Staining of Vascular Markers

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The antibodies against VEGFR3 (goat anti-mouse, R&D Systems (Minneapolis, MN) #AF743) and VEGFR2 (goat anti-mouse, R&D Systems #AF644) were used at a 1:50 dilution; the anti-LYVE (rabbit anti-mouse Abcam,Cambridge, MA) #ab14917) was used at 1:250; the anti-αSMA (mouse anti-mouse Dako,Santa Clara, CA) #M0851) was used at 1:100, and anti-PECAM/CD31 antibody (rat monoclonal, catalog #DIA-310; Dianova, Hamburg, Germany) was used at 1:250. Secondary antibodies were Donkey anti-rabbit 594 (#21207 ThermoFisher, Grand Island, NY); Donkey anti-goat 488 (Abcam #ab150129); Donkey anti-goat 546 (ThermoFisher #A11056); and Donkey anti-mouse 488 (Abcam #ab150105) were all used at a dilution of 1:250. Sections from a total of 3 pairs of A10ΔEC mice and littermate controls were analyzed per antibody, with at least one pair of littermates of each gender.
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2

Antibody Panel for Tumor Characterization

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Anti-Hpa2 monoclonal (20c5, 1c7) and polyclonal (#58) antibodies have been described previously [2 (link), 22 (link), 57 (link)]. Anti-phospho AMPK, anti-phospho-acetyl-CoA carboxylase (ACC), anti-phospho-STAT1, anti-phospho-Akt, and anti-major histocompatibility complex class I (MHC-I) antibodies were purchased from Cell Signaling (Boston, MA, USA). Anti-LYVE, anti-carbonic anhydrase 9 (CAIX), anti-Ki67, anti-collagen IV, anti-fibronectin, anti-Twist, anti-FSP1/S100A4, and anti-E-cadherin antibodies were purchased from Abcam (Cambridge, UK); Anti-syndecan-1 antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Anti-CD31 (PECAM) was purchased from Dianova (Hamburg, Germany). Rat anti-mouse F4/80 antibody was purchased from Serotec (Hercules, CA, USA). Anti-alpha-smooth-muscle actin (αSMA) antibody and poly(2-hydroxyethyl methacrylate) (poly-HEMA) were purchased from Sigma (St. Louis, MO, USA). Annexin V-FITC apoptosis detection kit was purchased from R&D systems (Minneapolis, MN, USA). Breast carcinoma tissue array (BR1505b) was purchased from US Biomax (Rockville, MD, USA).
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