Accesion numbers & Other nomenclature: AAA36539; Protein kinase B, v-AKT8, RAC-alpha Ser/Thr kinase like, STKc_PKB

Cell signaling mechanisms often transmit information via posttranslational protein modifications, most importantly reversible protein phosphorylation mediated by protein kinases.   AKT (also called protein kinase B) is a serine-threonine kinase that is activated downstream of growth factor receptors. It is a key player in signaling pathways that regulate energy metabolism, proliferation, and cell survival (1).  AKT1 was first described as the cellular homologue of the product of v-akt oncogene.  Protein kinase B, also known as Akt was first described as the cellular homologue of the product of the v-akt oncogene.  PKB comprise an N-terminal pleckstrin homology (PH) domain, a flexible hinge between the PH and the kinase domain, a catalytic (kinase) domain, and a C-terminal regulatory part (containing a hydrophobic motif, or HM).  Activation of AKT depends upon the integrity of PH homology domain, which mediates its membrane translocation and the phosphorylation of Thr308 in the activation loop and Ser473 (2).    AKT is a direct substrate of protein tyrosine kinase 6, that is not expressed in normal cells but induced in mammary cancer cells, phosphorylates AKT at Tyrosine residues Y315 and Y326.  

Protein kinase B is expressed in three isoforms (PKBalpha/AKT1, PKBbeta/AKT2 and PKBgamma/AKT3), AKT1 is expressed in multiple spliced variant forms.  The AKT1  gene is catalytically yin active but selectively activated by PDGF and the activation is abrogated by mutations in the PH domain.  The activation involves the IP3 kinase.  Understanding the regulation of Akt has been of major interest for elucidating the control of normal cellular physiology as well as malignant transformation. The activation of Akt involves phosphatidylinositol 3-kinase-dependent membrane localization followed by activating phosphorylation of Thr-308 and Ser-473. Many of the activating signals for Akt involve the stimulation of receptor and non-receptor tyrosine kinases, and the most potent activator known is the tyrosine phosphatase inhibitor pervanadate, highlighting a possible role for tyrosine phosphorylation in the regulation of the enzyme (3).  PKB is a 60 kDa protein expressed in majority of cells, mutations in this gene lead to proteus syndrome.  .

The AKT1-selective antibodies were generated against unique antigenic sequence form with in the C-terminal 50 amino acids.  The sequence was conserved in rat, mouse, human and many other species, but was unique only toAKT1 and was not present in other protein kinases.  The AKT antibodies were affinity purified over immobilized antigen based chromatography, and the affinity purified immunoglobulins are stabilized in antibody stabilization buffer.  FabGennix Int. Inc., will also provide limited quantities of antigenic blocking peptide for AKT1 antibodies.  FabGennix has a vast collection of antibodies to ser/threo, tyrosine kinase and related targets.  For a complete list of antibodies please visit www.FabGennix.com.  FabGennix Inc. will conjugate this and all its catalog antibodies with secondary enzymes (alk-Pase or HRP) or fluorescent probes upon request at a nominal cost.  FabGennix Int. Inc., will also provide and antigenic blocking peptide and western blot positive controls for it antibodies in ready-to-use buffer. 

Immunogen:        The antigenic peptide for protein kinase B antibody was chosen form with in the C-terminal last 50 aa.  The AKT peptide was post-synthetically modified  and conjugated to a carrier protein before injecting into host animals for antibody production.     

Concentration:     AKT-101AP and AKT-FITC antibodies concentration was in the range of 0.58-0.62mg/ml in antibody stabilization buffer. 

Applications:      Antibody AKT-101AP ideal for WB applications, use in IHC and IMM is not determined.  The dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions for specific assay.  WB;  1:500; IMM & i.p pull-down assays:> n.d.  n.d does not necessary means antibody will not work in that applications.  Investigators using this antibody in applications not listed here can request for a complimentary sample.             

Reactivity:         This antibody detects a band at 59kDa AKT (Protein kinase B) protein in WB blot positive controls and in several cell lines and tissues.   

Protocols:              Standard protocol for various applications (WB, IMM and IHC) of this antibody can be obtained upon request form technical services is provided with the product specification sheet, however, FabGennix Int. Inc.

Form/Storage:     The antiserum is supplied in antibody stabilization buffer with 0.02% sodium azide.  For long-term storage of antibodies, store at -20^oC.  FabGennix Int. Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DilUbuffer).  Working solutions of antibodies in DilUbuffer should be filtered through 0.45m filter after every use for long-term storage.

 

For research use only, not for diagnostic or therapeutic use.