Technical Data
A1124-05
Akt (Ser473) (Rac PKa, PKBa) BioAssay™ Phosphorylation Detection Kit
1Kit
Kits and Assays Storage: -20°CShipping: Blue Ice
Akt, also referred to as PKB and RAC, is the cellular homologue of the viral oncogene v-Akt (1-3). This protein kinase is activated by insulin and various growth factors and functions in a pathway involving PI3 kinase as activation is sensitive to wortmanin (2,3). Recent evidence suggests that Akt functions to promote cell survival by actively inhibiting apoptosis, although the pathways involved have not yet been identified (1,4). Akt is activated by phospholipid binding and phosphorylation within the activation loop at Thr308 and within the C-terminus at Ser473 (5). Currently, the enzymes activating Akt as well as the downstream substrates are not well understood. The Akt (Ser473) Antibody Kit provides reagents and protocols for the rapid analysis of Akt phosphorylation/activity. The Kit includes a phosphospecific antibody that recognizes Akt only when phosphorylated at Ser473, a control Akt antibody (phosphorylation-state independent), protein controls and protein markers for Western blots, and our HRP Western Detection System.

Applications:
Suitable for use in Western Blotting, Immunoprecipitation. Other applications not tested.

Recommended Dilution:
Western blotting: 1:1000 1:1000
Immunoprecipitation: 1:100 1:100

Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Purity: Purified by protein A and peptide affinity chromatography.
Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, BSA, 50% glycerol.
Specificity: Detects phosphorylation only at the Ser473 site and does not crossreact with related family members such as PKC or p70 S6 kinase.

Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
1. Franke, T.F. (1997) Cell 88, 435–437. 2. Burgering, B. T. and Coffer, P.J. (1995) Nature 376, 599–602. 3. Franke, T.F. et al. (1995) Cell 81, 727–736. 4. Kennedy, S.G. et al. (1997) Genes Dev. 11, 701–713. 5. Alessi, D.R. et al. (1996) EMBO J. 15(23), 6541–6551.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.