Immunofluorescence analysis of AKT3 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Akt3 (66C1247.1) Mouse Monoclonal Antibody (MA141201) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Storage buffer||PBS with 0.05% BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1:100-1:2000|
* 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.
The sequence is identical in human, mouse, rat, cow, dog, and chicken.
Suggested positive control: human kidney protein, antigen standard for AKT3 (transient overexpression lysate).
Akt, protein kinase B (PKB), is a serine/threonine kinase which is involved in many cellular signaling pathways and acts as a transducer of many functions initiated by growth factor receptors that activat phosphtidylinositol 3-kinase (PI 3-kinase). The major activity of Akt/PKB is to mediate cell survival. Akt/PKB is also believed to be a critical factor in the genesis of cancer as the tumor suppressor PTEN was found to antagonise PI-3 kinase and Akt/PKB kinase activity. There are 3 known isoforms of this enzyme in mammalian cells (1/alpha, 2/beta and 3/gamma). Akt1, 2 and 3 contain a key regulatory serine phosphorylation site in the carboxy-terminal region of the protein. Akt/PKB phosphorylation is critical for it activity.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.