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Our Abpromise guarantee covers the use of ab20654 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 96 kDa (predicted molecular weight: 96 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||1/2500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ICC/IF image of ab20654 stained rat PC12 cells. The cells were PFA fixed (10 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab20654, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Image courtesy of Human Protein Atlas
ab20654 staining HIF-2-alpha in Adrenal gland. The paraffin embedded human tissue was incubated with ab20654(1:2500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab20654 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.
Further results for this antibody can be found at www.proteinatlas.org