Anti-FOXG1 antibody

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXG1 antibody (AB18259)

IHC image of FoxG1 staining in mouse brain E14 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 ab18259, 0.5 μg/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.

Staining revealed in telencephalon (but not diencephalon) as expected.

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.

• IHC-P

AbReview4352****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXG1 antibody (AB18259)

ab18259 staining Foxg1 (1/50) in the telencephalon (but not the diencephalon) as expected. DAB-immunohistochemistry was performed on embryonic (E13.5) mouse brain (coronal paraffin-embedded sections) after microwave treatment with 10mM sodium citrate.

This image is courtesy of Vassiliki Fotaki, University of Edinburgh

• WB

Ap28169****

Western blot - Anti-FOXG1 antibody (AB18259)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab18259 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-FOXG1 antibody (ab18259) at 1 µg/mL

Lane 1:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lane 2:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 3:

E10 Mouse Embryo Brain Tissue Lysate at 10 µg

Lane 4:

Brain (Rat) Tissue Lysate at 10 µg

Lane 5:

Human spleen tissue lysate - total protein (negative control) at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) at 1/50000 dilution

Predicted band size: 52 kDa

Observed band size: 155 kDa,50 kDa,52 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-FOXG1 antibody (AB18259)

ab18259 detects a band of ~50kDa in brain lysates. This band is blocked by addition of the immunizing peptide ab19644 which suggests that is specific for FOXG1.

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab18259 overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP secondary antibody (ab97051), and visualised using ECL development solution.

All lanes:

Western blot - Anti-FOXG1 antibody (ab18259) at 1 µg/mL

Lane 1:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lane 2:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 3:

Brain (Rat) Tissue Lysate at 10 µg

Lane 4:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg with Human FOXG1 peptide (<a href='/en-us/products/unavailable/human-foxg1-peptide-ab19644'>ab19644</a>)

Lane 5:

Brain (Mouse) Tissue Lysate at 10 µg with Human FOXG1 peptide (<a href='/en-us/products/unavailable/human-foxg1-peptide-ab19644'>ab19644</a>)

Lane 6:

Brain (Rat) Tissue Lysate at 10 µg with Human FOXG1 peptide (<a href='/en-us/products/unavailable/human-foxg1-peptide-ab19644'>ab19644</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 52 kDa

Observed band size: 50 kDa

true

Exposure time: 1min