Immunofluorescent analysis of PAX8 (green) in 293T (left panel) and COS7 cells (right panel). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a PAX8 polyclonal antibody (Product # PA1-112) at a dilution of 1:400 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Non-human primate|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified internal fragment of human recombinant PAX8 expressed in E. coli|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:500|
|Immunoprecipitation (IP)||2-5 µg|
|Western Blot (WB)||1:500-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Western blot analysis of PA1-112 detects a prominent ~48 kDa protein in human adenocarcinoma SKOV-3 and SV40 transformed African green monkey (Cercopithecus aethiops) kidney cells. In SKOV-3 cells, 2 additional unknown bands at (~20 kD) and at ~150 kDa are also detected. Specificity of the antibody was confirmed in HeLa and 293T cells overexpressing full lenght PAX8. The 48 kDa band is enriched in immunoprecipitations from Cos7 lysate, however additional unknown high molecular weight bands are also enriched and detected by PA1-112.
PAX8 is a member of the paired box (PAX) family of transcription factors. Members of this gene family typically contain a paired box domain, an octapeptide, and a paired-type homeodomain. Pax8 is expressed in the developing and adult thyroid and kidney. In the thyroid, PAX8 regulates thyroglobulin, thyroidperoxidase,and NIS (sodium/iodide symporter) expression. Mutations in PAX8 have been associated with thyroid dysgenesis, thyroid follicular carcinomas and atypical follicular thyroid adenomas.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.