2015 top banner
 

Pharmacodynamic Assay for Profiling AKT Protein Activation Status


Pharmacodynamic Assay for Profiling AKT Protein Activation Status


PI3K/AKT/mTOR pathway is a central node in cell signaling downstream of growth factors, cytokines, and other cellular stimuli important in apotopsis and hence cancer. In humans, there are  three highly homologous members known as Akt1 (Uniprot P31749), Akt2 (Uniprot  P31751), and Akt3 (Uniprot  Q9Y243). It has been well-documented that  this pathway is overactive in a number of cancers.  Akt is activated by phosphorylation at the Ser473 and Thr308 positions by mTORC2 and PDK1 kinases, respectively. Faulty activation of Akt underlies the pathophysiological properties of a variety of complex diseases, including type-2 diabetes and cancer.  Constitutively activated Akt promotes cellular survival as well as resistance to treatment with chemotherapy and/or radiation therapy. In fact, several research groups are developing compounds targeting Akt as anticancer drugs.  Quantitation of the expression  of Akt proteins and their activated status  in response to the pathway inhibitors would have  great potential application in monitoring therapeutic efficacy.  In part, thanks to the funding support from the National Cancer Institute (NCI Awards HHSN26100900070C) Rockland Immunochemicals has developed  a suite of reagents and for  measuring phosphorylation of threonine residue pT308 and serine residue pS473 of Akt applicable to monitoring the Akt responses in the presence of Akt pathway inhibitors. 

 

 

AKT Antibody

 

 

Akt phospho pT308 Antibody

 

 

Akt phospho S473 Antibody

 

Localization of AKT (Green staining)  by fluorescence microscopy using rabbit anti-AKT antibody. 

100-401-401

 

Localization of active AKT (red signal) in human brain cerebellum tissue using  Mouse anti-AKT pT308 antibody

200-301-269

 

Western Blot using A431 cells stimulated with EGF and Mouse Anti-AKTpS473 antibody

200-301-B19

 


 

Suggested Products for Common Assays


For sandwich ELISA (E) to capture total Akt protein, rabbit raised using a C-terminal  peptide and sheep pan-reactive antibody raised against an internal peptide  that recognize  both inactive and active forms in combination with HRP conjugated species-specific secondary  antibody  should be used.

For Western Blotting (WB) use primary antibodies to respective targets followed with host-specific secondary antibody HRP conjugates.  Use our Blocking Buffers to reduce background and to enhance sensitivity.
 
For Immunofluorescence Microscopy (IF) use biotin conjugated antibodies in combination with streptavidin fluorescein conjugate.  For Immunohistochemistry (IHC) biotin conjugated antibodies can be used with streptavidin peroxidase conjugate  in combination with MaxTag™ DAB tablets.

 


 

Rockland Best-selling AKT-specific primary antibodies 

Product Name

Catalog # 

Applicatoin

Size

 Anti-AKT (RABBIT) Antibody

100-401-401

ELISA, WB, IHC, IF, FC

200 µL

 Anti-AKT (SHEEP) Antibody

600-601-401

ELISA, WB, IHC, IF, FC

100 µg

 Anti-AKT pS473 (RABBIT) Antibody

600-401-268

ELISA, WB, IHC, FC

100 µg

 Anti-AKT pT308 (MOUSE) Monoclonal Antibody

200-301-269

ELISA, WB, IHC, FC

100 µg

 Anti-AKT pS473 (MOUSE) Monoclonal Antibody

200-301-268

ELISA, WB, IHC, IF, FC

100 µg

 Anti-AKT pS473 (MOUSE) Monoclonal Antibody

200-301-268S

ELISA, WB, IHC, IF, FC

25 µL

 Anti-AKT2 (RAT) Monoclonal Antibody

200-501-E71

ELISA, WB, IHC, FC

100 µg

 Anti-AKT3 (MOUSE) Monoclonal Antibody

200-301-E75

ELISA, WB, IHC, FC

100 µg

 Anti-AKT2 (RAT) Monoclonal Antibody

200-501-J35

ELISA, WB, IHC, FC

100 µg

   

 


 

Antibodies and Reagents for Profiling AKT Actvation Status


Abstract


Rockland has developed a suite of reagents to detect phosphorylated and inactive forms of AKT proteins. These reagents are applicable for profiling AKT protein expression and phosphorylated activation status allowing monitoring of the efficacy of AKT pathway anti-cancer drugs. 

Applications


Reagents for profiling expression and activation of AKT proteins.

Advantages


AKT is activated by phosphorylation at the Ser473 and Thr308 positions by mTORC2 and PDK1 kinases, respectively. Faulty activation of AKT underlies the pathophysiological properties of a variety of complex diseases, including type-2 diabetes and cancer.  Constitutively activated AKT promotes cellular survival as well as resistance to treatment with chemotherapy and/or radiation therapy. Developed antibodies enables to rapidly quantify AKT protein expression and their phosphorylated stages in cells upon exposure to anticancer drugs, thereby allowing preclinical evaluation the anti-cancer drug efficacy and potential resistance problems.
To discuss licensing or custom services opportunities please contact:

Dan Kenney
Director, Scientific Business Development
(484)-791-3823

 

Get a Quote Button

 

Research Grants button

 

 

AKT Pathway Button