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Synthetic peptide corresponding to Human FOXP3 aa 400-500 conjugated to Keyhole Limpet Haemocyanin (KLH).
(Peptide available as
Our Abpromise guarantee covers the use of ab10563 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use a concentration of 4 µg/ml. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.|
|WB||Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 47.2 kDa).|
|IHC-Fr||Use a concentration of 0.625 µg/ml.|
|ICC||Use a concentration of 2.5 - 5 µg/ml.|
|ICC/IF||Use at an assay dependent concentration.|
This antibody recognises a single clean band in a HEK293 cell lysate that overexpresses human FOXP3. The band is not seen in untransfected cells and is blocked by the immunising peptide.
Immunofluorescent imaging of human cells (U2OS) with ab10563 (5 ug/ml) confirms the specificity of this anti-FOXP3 antibody, which recognises nuclear signal only; with no background cytoplasmic or nucleolar staining.
IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour). Primary antibody used at 5ug/ml in 5% milk / 0.2% TWEEN for 1 hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees C. Nuclei were stained with Hoechst stain.
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 Licor blocking buffer before being incubated with ab10563. Antibody binding was detected using Goat anti Rabbit IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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