All lanes: I7661-76T9 (1:1000) Lane 1: Hela whole cell lysate Lane 2: 293 whole cell lysate Lane 3: Jurkat whole cell lysate Lane 4: H. brain whole cell lysate Lane 5: K562 whole cell lysate Lysates/proteins at 20ug/lane. Secondary goat anti-rabbit IgG, (H+L), Peroxidase conjugated at 1:10,000. Predicted band size: 64kD. Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemistry formalin-fixed and paraffin-embedded human hepatocarcinoma with I7661-76T9, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of I7661-76T9 for immunohistochemistry.

Immunohistochemical analysis of paraffin-embedded human brain tissue using I7661-76T9 performed on the Leica® BOND RXm. Tissue was fixed with formaldehyde at RT; antigen retrieval was by heat mediation with a EDTA buffer (pH 9). Samples were incubated with I7661-76T9 (1:1000) for 1hr at RT. A undiluted biotinylated CRF Anti-Polyvalent HRP polymer antibody was used as the secondary antibody.

Overlay histogram showing K562 cells stained with I7661-76T9 (green line). The cells were fixed with 2% paraformaldehyde (10mins) and then permeabilized with 90% methanol for 10mins. The cells were then incubated in 2% BSA to block non-specific protein-protein interactions followed by I7661-76T9 (1:25) for 60mins at 37ºC. The secondary antibody used was goat-anti-rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed (1:200) for 40mins at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1ug/1x10e6 cells) used under the same conditions. Acquisition of >10,000 events performed.