TABLE 1
FROM:
Expression profiles associated with aggressive behavior in Merkel cell carcinoma
María-Teresa Fernández-Figueras, Lluís Puig, Eva Musulén, Montserrat Gilaberte, Enrique Lerma, Sergio Serrano, Carlos Ferrándiz and Aurelio Ariza
BACK TO ARTICLETable 1. Antibodies, suppliers, dilutions, and techniques used for immunohistochemistry and parameters evaluated
| Antibody | Source | Dilution | Technique | Parameters evaluated |
|---|---|---|---|---|
| Anti-human chromogranin A, mouse monoclonal antibody | Dako, Glostrup, Denmark | 1:600 (EDTA) | Dako Autostainer | Percentagea |
| Synaptophysin, mouse monoclonal antibody | Biogenex, San Ramón, CA, USA | 1:100 | Dako Autostainer | Percentage and intensityb and percentage (low:  10% of cellshigh: >10% of cells) |
| CD56 (NCAM), mouse monoclonal antibody | Novocastra, Newcastle upon Tyne, UK | 1:50 | Dako Autostainer | Percentage and intensityb |
| Anti-human cytokeratin 20, mouse monoclonal antibody | Dako | 1:50 | Dako Autostainer | 0: <1% of cells 1: >1% of cells |
| p21protein, mouse monoclonal antibody | Santa Cruz Biotechnology, Santa Cruz, CA, USA | 1:30 | Dako Autostainer | Percentagea |
| p53 protein (clone DO7), mouse monoclonal antibody | Novocastra | 1:500 | Dako Autostainer | Percentagea |
| Anti-human Bcl-2 oncoprotein, mouse monoclonal antibody | Dako | 1:350 | Ventana, Nexes | Percentage and intensityb |
| Anti-human Bcl-6 protein, mouse monoclonal antibody | Dako | 1:30 | Ventana, Nexes | Percentage and intensityb |
| CD117 (c-kit), rabbit polyclonal antibody | Dako | 1:50 | Ventana, Nexes | Percentagea |
| PTEN (A2B1), mouse monoclonal antibody | Santa Cruz Biotechnology | 1:50 | Ventana, Nexes | Intensityc |
| NF-kappaB/p50 Ab-2, rabbit polyclonal antibody | Neomarkers, Fremont, USA | 1:40 | Ventana, Nexes | Percentage and intensityb evaluated in tumor and in estroma |
| Survivin (FL-142) SC-10811, rabbit polyclonal | Santa Cruz Biotechnology | 1:30 | Dako Autostainer | Percentage and intensityb |
| Phospho-p38 MAPK (Thr180/Tyr182) 28B10, mouse monoclonal antibody | Cell Signaling, Danvers, MA, USA | 1:40 | Ventana, Nexes | Percentage and intensityb |
| Prostaglandin E synthase (microsomal), rabbit polyclonal antibody | Cayman, Massy, France | 1:300 | Ventana, Nexes | Percentage and intensityb |
| Prostaglandin E synthase-2 (microsomal), rabbit polyclonal antibody | Cayman, Massy, France | 1:380 | Ventana, Nexes | Percentage and intensityb |
HIF-1 , mouse monoclonal antibody | BD Biosciences, San José, CA, USA | 1:20 (buffer+LSAB) | LSAB+System-HRP, Dako | Intensityc |
| Vascular endothelial growth factor (VEGF) epitope-specific, rabbit polyclonal antibody | MD, Granada, Spain | 1:20 | LSAB+System-HRP, Dako | 0: absent; 1: <50%; 2: >50% |
| FLK-1 (A-3), mouse monoclonal antibody | Santa Cruz | 1:200 | Ventana, Nexes | Percentage and intensityb |
| Matrix metalloproteinase 1, mouse monoclonal antibody | Novocastra | 1:10 | LSAB+System-HRP, Dako | Percentage and intensityb |
| Matrix metalloproteinase 2, mouse monoclonal antibody | Novocastra | 1:10 | LSAB+System-HRP, Dako | Percentage and intensityb |
| MMP3 (stromelysin-1) Ab-2 (clone SL-1 IID4), mouse monoclonal antibody | Neomarkers | Prediluted (buffer) | LSAB+System-HRP, Dako | Percentage and intensityb |
| MMP7 (matrilysin) Ab-1 (clone ID2), mouse monoclonal antibody | Neomarkers | Prediluted (buffer) | Ventana, Nexes | Percentage and intensityb |
| Matrix metalloproteinase 9, mouse monoclonal antibody | Novocastra | Prediluted (buffer) | Ventana, Nexes | Percentage and intensityb |
| MMP10–2 (stromelysin-2) Ab-2 (clone IVC5), mouse monoclonal antibody | Neomarkers | Prediluted (buffer) | LSAB+System-HRP, Dako | Percentage and intensityb |
| MMP10–5 (stromelysin-2) Ab-5, rabbit polyclonal antibody | Neomarkers | Prediluted (buffer) | Ventana, Nexes | Percentage and intensityb |
| MMP13 (collagenase-3) Ab-1 (clone VIIIA2), mouse monoclonal antibody | Neomarkers | Prediluted (buffer) | Ventana, Nexes | Percentage and intensityb |
| MMP14/MT1-MMP Ab1, rabbit polyclonal antibody | Neomarkers | Prediluted (buffer) | LSAB+System-HRP, Dako | Percentage and intensityb |
| MMP15/MT2-MMP Ab1, rabbit polyclonal antibody | Neomarkers | Prediluted (buffer) | LSAB+System-HRP, Dako | Percentage and intensityb |
| MMP16/MT3-MMP Ab1, rabbit polyclonal antibody | Neomarkers | Prediluted (buffer) | LSAB+System-HRP, Dako | Percentage and intensityb |
| Matrix metalloproteinase 19, mouse monoclonal antibody | Novocastra | 1:10 | LSAB+System-HRP, Dako | Percentage and intensityb |
| Matrix metalloproteinase 23, mouse monoclonal antibody | Novocastra | Prediluted (buffer) | LSAB+System-HRP, Dako | Percentage and intensityb |
| TIMP1 Ab-7 (clone 2A5), mouse monoclonal antibody | Neomarkers | Prediluted (buffer) | LSAB+System-HRP, Dako | Percentage and intensityb |
| TIMP2 Ab-5 (clone 3A4), mouse monoclonal antibody | Neomarkers | Prediluted (buffer) | Ventana, Nexes | Percentage and intensityb |
| TIMP3 Ab-1, rabbit polyclonal antibody | Neomarkers | Prediluted (buffer) | Ventana, Nexes | Percentage and intensityb |
| TIMP4 Ab-1, rabbit polyclonal antibody | Neomarkers | Prediluted (buffer) | Ventana, Nexes | Percentage and intensityb |
 -Catenin (E-5), mouse monoclonal | Santa Cruz Biotechnology | 1:100 | Dako Autostainer | Percentagea |
| Anti-human CD44 hematopoietic, mouse monoclonal antibody | R&D Systems Europe, Abingdon, UK | 1:1000 | Ventana, Nexes | Percentage and intensityb |
| Anti-human CD44 variant 3, mouse monoclonal antibody | R&D Systems Europe | 1:600 | Ventana, Nexes | Percentage and intensityb |
| Anti-human CD44 variant 6, mouse monoclonal antibody | R&D Systems Europe | 1:1000 | Ventana, Nexes | Percentage and intensityb |
| Purified anti-MLH-1, mouse monoclonal antibody | BD Biosciences, San José, CA, USA | 1:50 | Dako Autostainer | 0: Total absence of nuclear staining 1: Presence of staining |
| Purified anti-MSH-2, mouse monoclonal antibody | BD Pharmingen | 1:200 | Dako Autostainer | 0: Total absence of nuclear staining 1: Presence of staining |
| MSH6/GTBP, IgG1 mouse monoclonal antibody | BD Biosciences, San José, CA, USA | 1:100 | Dako Autostainer | 0: Total absence of nuclear staining 1: Presence of staining |
| Purified anti-PMS2, mouse monoclonal antibody | BD Pharmingen | 1:60 | Ventana, Nexes | 0: Total absence of nuclear staining 1: Presence of staining |
a Percentage: The number of positive cells was scored as follows: 0: <2%; 1: 2–24%; 2: 25–50%; and 3: >50%.
b Percentage and intensity: After evaluating separately the percentage of positive cellsa and the intensity of stainingc, the two values were added and the final values were scored as follows: 0: absent; 1: 2 or 3 (1+1, 2+1, or 1+2); 2: 4 (1+3, 3+1, or 2+2); and 3: 5 or 6 (2+3, 3+2, or 3+3).
c Intensity: The degree of intensity was evaluated as follows: 1: faint; 2: moderate; and 3: intense.
