IHC image of FOXP2 staining in mouse e17 foetal brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16046, 0.1µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

ICC/IF image of ab16046 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16046, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

ab16046 at 1/1000 detecting FOXP2 from human 293T cell lysate (whole cell) (60ug/lane) by Western Blot. An HRP conjugated goat anti-rabbit IgG was used as the secondary and ECL was used as the detection method (1 minute exposure).

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Mouse spinal cord was fixed in paraformaldehyde, blocked in 1% BSA for 30 minutes then incubated with ab16046 at 1/8000 dilution for 18 hours. This image was submitted as part of a review by Jeremy Dasen.

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Image courtesy of Human Protein Atlas

ab16046 staining FOXP2 in human testis. Paraffin embedded human testis tissue was incubated with ab16046 (1/600 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab16046 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

Further results for this antibody can be found at www.proteinatlas.org

Anti-FOXP2 antibody (ab16046) at 1 µg/ml + HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Observed band size: 90 kDa why is the actual band size different from the predicted?
Additional bands at: 56 kDa, 70 kDa. We are unsure as to the identity of these extra bands.