Cd34 qbend 10

For immunohistochemistry, 2 μm thick sections were cut and antigens retrieved in an epitope retrieval solution of pH 8 (RE7116; Novocastra, Newcastle, UK) at 68 °C for 17 h in a stirred water bath. The antibody clones, dilutions and sources are as follows: CD99 (12E7, 1:25; Dako, Ely, UK), vimentin (V9, 1:100, Novocastra), CD31 (JC70, 1:100, Dako), CD34 (QBend10, 1:50, Dako), cytokeratin AE1/AE3 (1:100, Dako), CD45 (2B11+PD7/26, Dako), cytokeratin MNF116 (1:50, Dako), desmin (D33, 1:100, Dako), α-smooth muscle actin (SMA) (1A4, 1:200, Dako), epithelial membrane antigen (EMA) (E29, 1:100, Dako), HMB-45 (1:200, Dako), S100 (NCL-L-S100p, 1:1000, Novocastra), WT1 (C-19, 1:500, Santa Cruz, Insight Biotechnology Limited, PO Box 520, Wembley, Middlesex,UK), TLE1 (M-101, 1:50, Santa Cruz), ERG (Erg-1/2/3 C-1; 1:50, Santa Cruz), INI1 (1:25, BD Transduction Laboratories (BD Biosciences), Becton Dickinson UK, Oxford, UK), BCOR (C-10, 1:50, Santa Cruz), ETV4 (16, 1:50, Santa Cruz) and Ki-67 (MIB1, 1:200, Dako).

Sections were cut at 4 μm thick from formalin-fixed-paraffin-embedded (FFPE) tissue and immunostained using a VentanaBenchmark XT automatic stainer (Ventana, Tuscon, AZ, USA). Signals were revealed using the ultraView Universal Dab Detection kit (Ventana). The following antibodies were used: CD99 (12E7, Dako), EMA (E29, 1:50, Dako), desmin (D33, 1:80, Dako), cytokeratin AE1/AE3 (AE1/AE3, 1:50, Dako), caldesmon (h-CD, 1:100, Dako), myogenin (F5D, 1:100, Dako), S100 protein (Z0311, 1:800, Dako), CD34 (QBend-10, 1:25, Dako), INI1 (25, 1:50, BD Transduction Laboratories), BCOR (C-10, 1:50, Santa Cruz), ETV4 (16, 1:50, Santa Cruz). Immunohistochemistry for NTRK1 was performed using a 32 min incubation with the NTRK1 antibody (ab76291, clot EP1058Y, Abcam, dilution 1:200) on a Ventana ULTRA machine with Cell Conditioning Solution 1 pre-treatment for 64 min.