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Our Abpromise guarantee covers the use of ab6050 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 a concentration of 1 µg/ml. See Abreview.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 170 kDa. PubMed: 23293081|
Western blot for Gli3 antibody (ab6050) at 1/500 tested on the following tissue lysates:
Lane 1 : Human Brain
Lane 2: Human Lung
Lane 3: Human Spleen
Lane 4: Mouse Brain
Lane 5: Mouse Lung
Secondary ab: Goat anti-rabbit IgG ab6721 (1/5000)
Exposure time: 3 minutes
Expected molecular weight: 2 isoforms of Gli3 exist, one is the full length 170-190kDa and the other is a truncated isoform at ~80kDa.
Cell lysates were loaded at 20
NB: This image shows the truncated isoform of Gli3 is detected by ab6050 in human lung (we do not know the identity of the 120-130kDa band). It was not possible to detect Gli3 immunoreactivity by WB in the other lysates tested. This is likely due to low expression/abundance of Gli3 in these tissue lysates. Furthermore, Gli3 expression is likely to be developmentally regulated and induced, making it difficult to succesfully detect in whole tissue homogenates. The staining on the WB shows low background and minimal cross reactivity in whole homogenates. The 50kDa band is likely due to non-specific binding of secondary antibody.
ab6050 at 0.625 µg/ml staining human glioblastoma.