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Our Abpromise guarantee covers the use of ab18259 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).|
|ChIP||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
ab18259 staining FOXG1 in Rat embryonic cortex cells overexpressing FOXG1 by ICC/IF (Immunocytochemistry/ immunofluorescence). Cells were fixed with formaldehyde and blocked with 1o% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/500) for 24 hours at 4°C. A Cy3®-conjugated anti-rabbit IgG polyclonal was used as the secondary antibody (1/500).
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.
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, and visualised using ECL development solution.
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.
ab18259 staining FOXG1 in human A549 cells by immunocytochemistry/immunofluorescence.Cells were fixed in paraformaldehyde, permeabilized using 0.25% Triton, blocked with 1% BSA for 1 hour at room temperature and then incubated with ab18259 at a 1/400 dilution for 1 hour. The secondary used was an Alexa-Fluor 488 conjugated goat polyclonal used at a 1/250 dilution.
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