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Our Abpromise guarantee covers the use of ab10648 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/2000. Predicted molecular weight: 110 kDa. 1/2000 this dilution is determined by blotting using a whole extract of transfected cells expressing recombinant human FGFR2. Predicted molecular weight: 110 kDa.|
|IP||1/1000. 1/1000, this dilution is determined by immunoprecipitation using a whole lysate of transfected cells expressing recombinant human FGFR2.|
|IHC-P||1/1000. Perform enzymatic antigen retrieval before commencing with IHC staining protocol. The recommended enzyme is trypsin.|
ab10648 staining FGFR2 in the epithelium (top) and transition zone (bottom) of Embryonic Mouse eye tissue sections by Immunohistochemistry ((IHC) paraffin-embedded sections). Tissue was fixed, embedded in paraffin and sectioned. Sections were trypsinized for 40 minutes at room temperature in a humidified chamber, washed in PBS, and then incubated for 1 hour with a 0.5% Triton X-100 and 0.3 M glycine in PBS in a humidified chamber. After antigen retrieval, sections were blocked in 0.5% nonfat dry milk, 10% horse serum, and 0.2% Triton X-100 diluted in PBS for 3 hours at room temperature. An Alexa Fluor®568-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded analysis of hb/hb and control mouse embryo (P5) tissue sections labelling FGFR2 with ab10648. Samples were fixed overnight at 4°C in 10% neutral-buffered formalin, embedded in paraffin and cut into 8µm sections.
At this stage, FGFR2 is located in hair cells (black arrowheads) and tectorial membrane (red arrowhead). No significant differences in the Fgfr2 protein levels are detected in hb/hb mutants compared to controls. Scale bar: 10µm.