Clone uj127

Immunohistochemistry was performed as described earlier. IHC for detection of L1CAM (mouse monoclonal antibody, clone UJ127.11; Sigma Aldrich, St. Louis, MO, USA; 1 : 1500), IDH1-R132H (clone H09, 1 : 50; Dianova, Hamburg, Germany), ATRX (1 : 500; Sigma-Aldrich, St. Louis, MO, USA), P53 (1 : 100; Dako, Carpinteria, CA) and Ki67 (1 : 100; Dako, Carpinteria, CA) was performed on an automated BenchMark Ultra (Ventana Medical systems, Roche, SW).
Immunohistochemical evaluation was independently conducted by two pathologists blinded for patient characteristics and outcome, with discrepancies resolved by consensus under a microscope for multi-viewing.
The result of positive L1CAM staining was used the adjusted Allred scoring system to evaluate the results of L1CAM expression, and the total value was 0-12 by positive ratio × staining intensity. On account of normal brain tissues can weakly express L1CAM, diffuse and strong staining localized in tumor cell membrane and cytoplasm was defined as highly positive. Normal kidney tissue was used as a positive control and vascular endothelial cell was an internal negative control.

Unstained 3-μm-thick slides of formalin-fixed, paraffin-embedded (FFPE) tissues were obtained and submitted for immunostaining using an automated stainer (Dako Omnis, Glostrup, Denmark). The following primary antibodies were used: Glial Fibrillary Acidic Protein (GFAP) (1:200, clone 6F2, Dako, Glostrup, Denmark), Olig2 (1:500, clone OLIG2, Sigma-Aldrich, Saint-Louis, USA), SOX10 (1:50, clone A-2, Diagomics, Blagnac, France), neurofilament (1:100, clone NF70, Dako, Glostrup, Denmark), NeuN (1:1000, clone A60, Sigma-Aldrich, Saint-Louis, USA), synaptophysin (1:150, clone Synap, Dako, Glostrup, Denmark), chromogranin A (1:200, clone LK2 H10, Diagnostic Biosystem, Pleasanton, USA), EMA (1:200, clone GM008, Dako, Glostrup, Denmark), L1CAM (1:500, clone UJ127.11, Sigma-Aldrich, Saint-Louis, USA), NFκB (1:6000, clone D14E12, Cell Signaling Technology, Danvers, USA), H3K27me3 (1:2500, polyclonal, Diagenode, Liege, Belgium), and Ki-67 (1:200, clone MIB-1, Dako, Glostrup, Denmark). External positive and negative controls were used for all antibodies and staining. MIB-1 labeling index was jointly estimated by two neuropathologists in a hot-spot area.

Unstained 3-μm-thick slides of formalin-fixed, paraffin-embedded tissues were obtained and submitted for immunostaining with an automated stainer (Dako Omnis, Glostrup, Denmark). The following primary antibodies were used: CD56 (pre-diluted, clone 123C3, Dako, Glostrup, Denmark), Glial Fibrillary Acidic Protein (GFAP) (1:200, clone 6F2, Dako, Glostrup, Denmark), Olig2 (1:500, clone OLIG2, Sigma-Aldrich, Saint-Louis, USA), vimentin (1:800, clone V9, Dako, Glostrup, Denmark), neurofilament (1:100, clone NF70, Dako, Glostrup, Denmark), NeuN (1:1000, clone A60, Sigma-Aldrich, Saint-Louis, USA), synaptophysin (1:150, clone Synap, Dako, Glostrup, Denmark), EMA (1:200, clone GM008, Dako, Glostrup, Denmark), CK18 (1:200, clone 6F2, Dako, Glostrup, Denmark), smooth muscle actin (1:4000, clone 1A4, Dako, Glostrup, Denmark), NFκB (1:6000, clone D14E12, Cell Signaling Technology, Danvers, USA), L1CAM (1:500, clone UJ127.11, Sigma-Aldrich, Saint-Louis, USA), and Ki-67 (1:200, clone MIB-1, Dako, Glostrup, Denmark). Reticulin staining was performed using the Reticulin silver plating kit according to Gordon & Sweets (Merck Millipore, Guyancourt, France). External positive and negative controls were used for all antibodies and staining.