Western blot analysis was performed on whole cell extracts (30 ug lysate) of HeLa (Lane 1), MDA-MB-231 (Lane 2), HEK-293 (Lane 3), A549 (Lane 4), U-87 MG (Lane 5), T98G (Lane 6), Jurkat (Lane 7) and Caco-2 (Lane 8).The blots were probed with AKT2/AKT3 Rabbit polyclonal Antibody (Product# PA1-41144, 2 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution). A 56 kDa band corresponding to AKT2/AKT3 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues 319-331 of human AKT2.|
|Contains||0.02% 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 *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Western Blot (WB)||1-3 µg/ml|
* 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.
Suggested positive control: HeLa, antigen standard for AKT2 (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 activate phosphatidylinositol 3-kinase (PI 3-kinase). Akt2 is amplified and overexpressed in some human cancers.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.