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Our Abpromise guarantee covers the use of ab5073 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.|
|IHC-P||Use a concentration of 4 - 8 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
|WB||Use a concentration of 0.5 µg/ml. Detects a band of approximately 80-85 kDa (predicted molecular weight: 85 kDa).Can be blocked with Human STAT3 peptide (ab23064).|
|ELISA||Use at an assay dependent dilution.
Immunofluorescence detection using Goat polyclonal to STAT3 (ab5073) on rat (mixed neuron/glia) cell culture. Cells were fixed with paraformaldehyde, no antigen retrieval step was necessary. Primary antibody ab5073 was incubated for 1hr at room temperature (25 µg/ml). The confocal image shows anti-STAT3 positive immunoreactivity (red) in glial processes and nuclei. Blue nuclear counterstain is shown. Secondary antibody was donkey anti-goat IgG Alex fluor 568 (1/800) ab175474).
ab5073 (4µg/ml) staining of paraffin embedded Mouse Thymus shows preferential staining of the medula over the cortex. Steamed antigen retrieval with Tris/EDTA buffer pH 9, HRP-staining.
Immunofluorescence detection of STAT3 using Goat polyclonal to STAT3 (ab5073) on human HEK cell culture. 5% BSA was used as a blocking agent for 40 mins. Cells were fixed with methanol; no antigen retrieval step was necessary. Primary antibody ab5073 was incubated for 12 hour at RT (1/100). The confocal image shows anti-STAT3 cytosolic and plasma membrane localisation as expected.
ab5073 at 10ug/ml staining STAT3 in mouse thymus tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). The section incubated with biotinylated mouse anti goat as secondary, then Streptavidin-AP conjugate and developed using BCIP/NBT detection system.
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