Rabbit polyclonal isotype control antibody

Formalin-fixed, paraffin-embedded (FFPE) sections from primary MCC tumors were prepared and analyzed as previously described (55 (link)). The primary antibodies were cytokeratin 20 (CK20) antibody (dilution 1:50 [Dako]), MCPyV LT antibody CM2B4 (dilution 1:125 [Santa Cruz Biotechnology]), and anti-stathmin 1 antibody (dilution 1:250 [Abcam]). An isotype-matched irrelevant antibody was used as a negative control on serial sections of tissues in parallel, Dako X0943 was used for the CK20 primary antibody, and the rabbit polyclonal isotype control antibody (Abcam) was used to match the stathmin primary antibody. Sections were incubated with appropriate secondary antibodies labeled with different fluorochromes: Alexa Fluor 488 IgG2B and Alexa Fluor 633 IgG2A [Invitrogen] and IgG (H+L)-tetramethyl rhodamine isocyanate (TRITC) (Jackson ImmunoResearch). Nuclear counterstaining was with bis-benzimide (Invitrogen). All slides were mounted with Immu-Mount, and images were captured with a Zeiss LSM 510 confocal microscope (56 (link)).

Formalin-fixed, paraffin-embedded (FFPE) sections from primary MCC tumours were purchased from Origene and analysed as previously described [32 (link)]. Primary antibodies were: FITC-conjugated anti-CK20 (Dako, dilution 1:50), MCPyV LT CM2B4 (Santa Cruz Biotechnology, dilution 1:125) and ADAM 10 and 17 (Abcam, dilution 1:250). An isotype-matched irrelevant antibody was used as a negative control on sections of tissues in parallel, a rabbit polyclonal isotype control antibody (Abcam) was used to match the ADAM 10 primary antibody. Sections were incubated with appropriate secondary antibodies labelled with different fluorochromes (Alexa Fluor 546 IgG2B, 643 IgG2A, Invitrogen, and IgG (H+L)-TRITC, Jackson ImmunoResearch). All slides were mounted with Immuno-Mount and images were captured with a Zeiss LSM880 confocal laser scanning microscope.

FFPE sections from primary MCC tumors were purchased from Origene and analyzed as previously described (83 (link)). The primary antibodies were CK20 (Dako; dilution, 1:50), MCPyV LTA CM2B4 (Santa Cruz Biotechnology; dilution, 1:125), and anti-cortactin (Abcam; dilution, 1:250). An isotype-matched irrelevant antibody was used as a negative control on sections of tissues in parallel, and a rabbit polyclonal isotype control antibody (Abcam) was used to match the cortactin primary antibody. The sections were incubated with appropriate secondary antibodies labeled with different fluorochromes [Alexa Fluor 488 IgG2B and 633 IgG2A (Invitrogen) and IgG(H+L)-tetramethyl rhodamine isocyanate (TRITC) (Jackson ImmunoResearch)]. All slides were mounted with Immuno-Mount, and images were captured with a Zeiss LSM 510 confocal microscope.