Immunofluorescent analysis of Phospho-GSK-3-alpha/beta pSer21/9 (green) in HeLa cells either left untreated (left panel) or treated with 100nM insulin for 20 minutes (right panel). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Phospho-GSK-3-alpha/beta pSer21/9 polyclonal antibody (Product # PA5-17899) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 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 ArrayScan or ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse, Rat, Zebrafish, Non-human primate|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide corresponding to the sequence of human GSK-3 alpha|
|Purification||Antigen affinity chromatography|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50|
|Western Blot (WB)||1:1000|
* 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.
Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
It is not recommended to aliquot this antibody.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Carbohydrate metabolism Other polysaccharide metabolism Glycogen metabolism Protein metabolism and modification Protein modification Protein phosphorylation Signal transduction Cell surface receptor mediated signal transduction Other receptor mediated signaling pathway Intracellular signaling cascade Developmental processes Embryogenesis Segment specification Ectoderm development Neurogenesis Mesoderm development Cell cycle Mitosis Cell proliferation and differentiation Other intracellular signaling cascade