Western blot analysis of TFAP2A over-expressed 293 cell line, cotransfected with TFAP2A Validated Chimera RNAi ( Cat # H00007020-R01V ) (Lane 2) or non-transfected control (Lane 1). Blot probed with TFAP2A monoclonal antibody (M01), clone 2G5 (Cat # H00007020-M01 ). GAPDH ( 36.1 kDa ) used as specificity and loading control.
Human HepG2; WB Suggested Anti-TFAP2A Antibody Titration: 1.25ug/ml. ELISA Titer: 1:312500. Positive Control: HepG2 cell lysate; TFAP2A antibody - C-terminal region (P100878_T100) in Human HepG2 cells using Western Blot
AP07641PU-N TFAP2A antibody staining of Formalin-Fixed Paraffin-Embedded Human Kidney at 2.5 µg/ml followed by biotinylated anti-Goat IgG secondary antibody, Alkaline Phosphatase-Streptavidin and Chromogen.
Western blot analysis of TFAP2A polyclonal antibody ( Cat # PAB3818 ) in NCI-H460 cell line lysates ( 35 µg/lane ) . TFAP2A ( arrow ) was detected using the purified polyclonal antibody ( 1 : 60 dilution ) .
Immunohistochemical analysis of AP2 alpha/beta staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Immunofluorescent analysis of AP2 alpha/beta staining in HepG2 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).
Anti-AP2 alpha/beta Antibody was used in an Electrophoretic Mobility Shift Assay (EMSA) to supershift the protein-DNA complex. Radiolabelled double-stranded DNA oligonucleotides (10.000 cpm per lane) harbouring a binding site for AP2 alpha/beta were incubated with each 2 ug of nuclear extract (NE) from HeLa and Caski cells respectively. Samples were incubated for 30 minutes at room temperature to allow the formation of protein-DNA complexes. Anti-AP2 alpha/beta Antibody were added to the samples (as indicated) and incubated for further 60 minutes at 4 C. Samples were separated in a 5.5% PAGE. The Gel was dried under vacuum and for autoradiography a X-ray film was exposed with an intensifying screen for 2 days at -80 C. Specific protein-DNA complexes were quantitatively supershifted with Anti-AP2 alpha/beta Antibody verifying the binding of AP2 alpha/beta to the DNA oligonucleotide.
ChIP analysis of Cervical cancer cell lines lysate incubated for 12 hours at 4 C. Cross-linking (X-ChIP) using formaldehyde for 10 minutes. Detection step: Semiquantitative PCR. Positive control: Tumor cell lines Hela. Negative control: Human primary keratinocytes.