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Our Abpromise guarantee covers the use of ab9758 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 at an assay dependent concentration. PubMed: 25421510|
|IHC-P||Use at an assay dependent concentration. PubMed: 25207642|
|WB||1/100 - 1/500.
In reduced samples, the expected bands are 12 kDa (mature protein) and 44 kDa (unprocessed protein). Expected band at 25 kDa in non-reduced samples (homodimer of mature protein). Glycosylations may also cause unprocessed protein to run a few kDa higher (around 50 kDa). TGFB1 also heterodimerizes with TGFB2, so there is potential for multiple different band sizes in WB.
Incubated with the primary antibody overnight at 4°C.
Incubated with the secondary antibody for 30 min at room temperature.
Five micron sections of paraffin embedded peritoneum tissue were mounted onto electrostatically charged microscope slides and dewaxed in xylene (2×5min) and rehydrated. Antigens were retrieval by pressure-cooking slides in 10 mM Tris 1 mM EDTA pH 9, for 5 mins. Slides were washed in H2O before incubation with 3% hydrogen peroxide for 30 min. Slides were blocked then incubated with ab9758 diluted 1/1000 or rabbit IgG in blocking buffer overnight at 4°C. Slides were washed in TBST20 and incubated for 30 min with secondary antibody and washed again before incubation with 3, 3′-diaminobenzidine for 5 min. Slides were then counterstained with hematoxylin, dehydrated and visualized by light microscopy, using an Olympus Provis microscope equipped with a Kodak DCS330 camera.
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