Anti-TTF1 antibody [EP1584Y] (ab76013)

Immunocytochemistry/ Immunofluorescence analysis of human thyroid cells labeling TTF1  with purified ab76013 at 1/500 dilution (0.3μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% tritonX-100. Antigen retrieval was performed using a heated citrate solution (10mM citrate PH 6.0 + 0.05% Tween-20). ab150077, a Goat anti rabbit IgG(Alexa Fluor^® 488) secondary antibody was used at 1/1000 dilution. PBS instead of the primary antibody was used as a control. DAPI nuclear staining.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling TTF1 with purified ab76013 at 1/250 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

Blocking and diluting buffer: 5% NFDM/TBST.

Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling TTF1 with purified ab76013 at 1/20 dilution (10 ug/ml). Cells were fixed with 4% paraformaldehyde. A Goat anti-rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black), Unlabeled control - cells without incubation with primary antibody and secondary antibody (Blue).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat lung tissue sections labeling TTF1 with purified ab76013 at 1/250 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

Blocking and diluting buffer: 5% NFDM/TBST.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse lung tissue sections labeling TTF1 with purified ab76013 at 1/250 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling TTF1 with unpurified ab76013 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

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Immunofluorescence analysis of rat bronchi cells, staining TTF1 with unpurified ab76013.

Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 27°C. Samples were incubated with primary antibody (1/500 in blocking buffer) for 1 hour at 16°C. An AlexaFluor®594-conjugated goat anti-rabbit polyclonal IgG (1/500) was used as the secondary antibody.

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Unpurified ab76013 staining TTF1 in murine fetal lung tissue by Immunohistochemistry (Frozen sections).Tissue was fixed in formaldehyde and permeabilized using PBST. Samples were then blocked using 1.5% serum for 20 minutes at 25°C, then incubated with ab76013 at a 1/400 dilution for 12 hours at 4°C. A biotin conjugated goat anti-rabbit IgG was used as the secondary at a 1/200 dilution.

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Overlay histogram showing A549 cells stained with unpurified ab76013 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76013, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling TTF1 with purified ab76013 at 1/250 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.