For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome
Synthetic peptide corresponding to Human AP2 alpha.
Database link: P05549
Our Abpromise guarantee covers the use of ab52222 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration.|
|ICC/IF||1/1000. (see Abreview)|
|EMSA||Use at an assay dependent concentration.|
|WB||1/500 - 1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).|
|IHC-P||Use a concentration of 5 µg/ml.|
|EMSA||Use at an assay dependent concentration. PubMed: 20671194|
Immunohistochemical analysis of formaldehyde-fixed paraffin-embedded murine embryonic tissue sections, labelling AP2 alpha with ab52222 at a dilution of 1/3000 incubated for 2 hours at 21°C in TBS, BSA and azide diluent. Heat mediated antigen retrival was with citric acid. Blocking was with 1% BSA incubated for 10 minutes at 21°C. Secondary was a goat anti-rabbit polyclonal biotin conjugate at 1/300. In this day 18 embryo, nuclear positivity was also detected in spinal cord, ganglia, kidney tubules and the keratinised portion of the stomach.
SiHa (Human cervical cell lines) cells stained for AP2 alpha (Green) using ab52222 (1/500 dilution) in ICC/IF. Secondary used is an Alexa-Fluor®488-conjugated Goat Anti-Rabbit IgG (H+L).
Antibody anti-AP-2 alpha (ab52222) was used in an Electrophoretic Mobility Shift Assay (EMSA) to supershift the protein-DNA complex.
Radiolabeled, double-stranded DNA oligonucleotides (10.000 cpm per lane) harbouring a binding site for AP-2 alpha were incubated with each 2 µg 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. 2 µg of anti-AP-2 alpha 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 for 30 minutes at 280 V and further 75 minutes at 350 V. 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 antibody anti-AP-2 alpha (ab52222), verifying the binding of AP-2 alpha to the DNA oligonucleotide.
IHC image of AP2 alpha staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab52222, 5 µg/ml, for 15 minutes 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"