The serine–threonine kinase Akt (also known as protein kinase B) is a central convergence node in a broadly influential signaling network. Akt activation serves as a master switch of these cellular signaling pathways, generating a multitude of intracellular responses through a plethora of downstream targets and interacting partners. Because of its pivotal role in cell signaling, and the consequences of that signaling in diseases ranging from cancer and diabetes to neurodegeneration, Akt is one of the most actively studied kinases in both basic research and drug development. Here we highlight our newest products for analyzing the Akt signaling pathway and its downstream targets.

Life Technologies offers Novex® antibodies, ELISAs, Luminex® multiplex assays and growth factors for key targets in the Akt signaling cascade. 

 

Find Akt signaling pathway primary antibodies, antibody pairs, ELISAs and Luminex® assays by protein target: 


Novex® Multiplex Panels

Targets A–E  Targets F–N  Targets O–V
     akt pathway

Key Akt Pathway Targets

Akt is expressed as three isoforms — AKT1/ PKBα, AKT2/ PKBβ and AKT3/ PKBγ, respectively (1). An amino terminal pleckstrin homology (PH) domain, a central serine–threonine catalytic domain, and a small carboxy‑terminal regulatory domain characterize all the three isoforms. The PH domain binds to phosphatidylinositol-3,4-bisphosphate (PIP2) and phosphatidylinositol-3,4,5-trisphosphate (PIP3), products of PI3K. This binding causes Akt to locate to the plasma membrane, where it becomes phosphorylated by phosphoinositide-dependent kinase 1 (PDK1) on Thr308 in the activation loop of the catalytic domain. This phosphorylation leads to activation. Full activation requires phosphorylation at a second site (Ser473).  Current evidence leads to the mTOR–rictor complex (mTORC2) as the primary kinase for the second phosphorylation event, although other kinases like Ilk (integrin linked kinase) (2), PDK1 (3), DNA-dependent protein kinase (DNA-PK) (4), ATM (ataxia telangiectasia mutated) have also been identified (5).

 

  1. Vivanco, I. & Sawyers, C. L. (2002) Nature Rev.Cancer 2, 489–501
  2. Persad S et.al. (2000) Proc Natl Acad Sci USA 97: 3207–3212
  3. Balendran A. et.al. (1999) Curr Biol 9: 393–404
  4. Feng et.al. (2004) J Biol Chem 279: 41189–41196
  5. Dong LQ, Liu F. (2005) Am J Physiol Endocrinol Metab 289: E187–E196

 

Western Blot Analysis

Western Blot with with Anti-AKT Antibody

Western blot analysis of Akt [pS473] ABfinity™ recombinant antibody.


Anti–AKT [pS473] ABfinity™ monoclonal antibody was used at 0.1 µg/mL to label AKT [pS473] in lysates of untreated 3T3 (lane 1) or PDGF-treated (lane 2) NIH/3T3 cells.

Indirect ELISA

Direct ELISA Akt Antibody

Indirect ELISA with anti–AKT [pT308] ABfinity™ recombinant rabbit monoclonal antibody.


Indirect ELISA was performed using various dilutions of anti–AKT [pT308] ABfinity™ monoclonal antibody to detect recombinant AKT [pT308] protein coated onto the plate.

Noxex® Multiplex Assays

Akt Multiplex Assays
(click to enlarge)

Multiplex quantitation of Akt pathway signaling proteins using the Luminex® platform..


MCF-7 cells were treated with insulin to induce protein phosphorylation, and then cell lysates were prepared. Protein concentrations in the MCF-7 cell lysates were measured for seven phosphorylated proteins—AKT [pS473], GSK-3β [pS9], IGF-1R [pYpY1135/1136], IR [pYpY1162/1163], IRS-1 [pS312], p70S6K [pTpS421/424], PRAS40 [pT246]—using the Akt Pathway Phospho Magnetic 7-Plex Panel and the Luminex® 200™ system.

Find Akt Protein Targets A–E

Target ABfinity™ Recombinant Monoclonal Antibody ABfinity™ Recombinant Oligoclonal Antibody Other Primary Antibodies** Antibody Pairs ELISA Proteins Luminex® Assays
4E–BP1  AHO1382
(Ms)

See all antibodies
 KHO0681  
4E–BP1 [pT37]700238      
4E–BP1 [pT46]700397 441170G
(Rb)
 KHO0691  
Akt 71000544609M
(Ms)

See all antibodies
 KHO0101
KHO0531
See all ELISAs
 LHO0091
(Singleplex)
Akt [pS473] & [npS473]700392
700256
 44622M [npS473]
(Ms)

44621G
(Rb)
CHO0115KHO0111
KHO0541
See all ELISAs
 LHO0101
(Singleplex)
Akt [pT308]70105271012244602G
(Rb)
 KHO0201  
Akt2  393900
(Ms)
    
ALK  180266
(Rb)

513900
(Rb)
    
Androgen Receptor  415900
(Ms)
    
Angiopoietin–2 (Angiogenin)  407600
(Rb)
 KHC1641  


Target ABfinity™ Recombinant Monoclonal Antibody ABfinity™ Recombinant Oligoclonal Antibody Other Primary Antibodies** Antibody Pairs ELISA Proteins Luminex® Assays
Angiopoietin 3 (N–term)  425500
(Rb)

425200
(Rb)
    
Angiopoietin 4 (N–term)  425300
(Rb)
    
Axl  A14027
(Rb)
    
BAD [pS128]  44523G
(Rb)
    
BMP2 710022   PHC7141
PHC7143
PHC7145
PHC7146
 
c–Kit (CD117)  348800
(Rb)

348800
(Rb)

See all antibodies
    
c–Kit [pY703] (CD117)  44492
(Rb)
    
c–Kit [pY705] (CD117) 700135      
c–Kit [pY719] (CD117)  A14013
(Rb)
    
c–Kit [pY721] (CD117)  44494G
(Rb)
    
c–Kit [pY730] (CD117)  44496G
(Rb)
    
c–Kit [pY823] (CD117)  44498G
(Rb)
    
c–Kit [pY936] (CD117)  44500G
(Rb)
    
c–Kit [pYpY568⁄570] (CD117)  44490G
(Rb)
    
c–Met700261718000182257
(Rb)

370100
(Ms)

See all antibodies
CHO0285KHO0251  
c–Met (soluble)   CHO0315KHO2031  
c–Met [pY1003]  44882G
(Rb)

A14006
(Rb)
    
c–Met [pYpYpY1230⁄1234⁄1235]700139 44888G
(Rb)
 KHO0281  
c–Met [pYpY1234⁄1235]  44887G
(Rb)
    
c–Met [pY1349]  44892G
(Rb)
    
C–Met [PY1365]  44896G
(Rb)
    
Catenin–β 712700AHO0462
(Rb)

138400
(Ms)

See all antibodies
 KHO1211  
Catenin–β [npaa 27–37]  44207M
(Ms)
    
Catenin–β [npaa 35–50]  44206M
(Ms)
    
Catenin–β [pS45]  44208G
(Rb)

A14041
(Rb)
    
Catenin–β [pY86]  44210M
(Ms)
    
Catenin–β⁄γ [pY654⁄pY643]  44212M
(Ms)
    
CD135  MHCD13504
(Ms)
    
Cyclin D1  134500
(Ms)

AHF0088
(Ms)

See all antibodies
    
EGF    KHG0062
KHG0061
See all ELISAs
PHG0311
PHG0313
PHG0314
PHG0315
LHG0061
(Singleplex)
EGFR700308
700994
 44799G
(Rb)

AHR5062
(Ms)

See all antibodies
 KHR9061
See all ELISAs
 LHR9061
(Singleplex)
EGFR [pY845]  44784G
(Rb)
    
EGFR [pY992]  44786G
(Rb)
    
EGFR [pY1068] 70105371012344788G
(Rb)
 KHR9081 LHR9081
(Singleplex)
EGFR [pY1086]  44790G
(Rb)

369700
(Rb)
    
EGFR [pY1148] 71021344792G
(Rb)
    
EGFR [pY1173]  44794G
(Rb)
 KHR9071  
EphA2 Receptor  347400
(Rb)

374400
(Ms)
    
EphA3 Receptor  348500
(Rb)

373200
(Ms)
    
EphA4 Receptor  347900
(Rb)

371600
(Ms)
    
EphB2 Receptor  366100
(Rb)

371700
(Ms)
    
EphB4 Receptor  352900
(Ms)

371800
(Ms)
    
ErbB–2 (HER–2)  280004
(Rb)

AHO1011
(Ms)

See all antibodies
 KHO0701  
ErbB–2 [pY1248] (HER–2)700635 44904G
(Rb)
    
**Gp=guinea pig monoclonal IgG antibody; Ms=mouse monoclonal IgG antibody; Rb=rabbit polyclonal IgG antibody.

Find Akt Protein Targets F–N

Target ABfinity™ Recombinant Monoclonal Antibody ABfinity™ Recombinant Oligoclonal Antibody Other Primary Antibodies** Antibody Pairs ELISA Proteins Luminex® Assays
FGF–basic  710148  KHG0021
(96 Tests)

KHG0022
(192 Tests)
13256029LHG0021
(Hu Singleplex)

LMG0021
(Ms Singleplex)
FGFR1  133100
(Ms)

3379200
(Ms)
    
FOXO3 [pS207]⁄FOXO1 [pS212]  441230G
(Rb)
    
GATA4 [pS105]  44948
(Rb)
    
GDF–15  421700
(Rb)
    
GSK–3β701102710132399500
(Ms)

See all antibodies
 KHO0451 LHO0451
(Singleplex)
GSK–3β [pS9]701069710100  KHO0461 LHO0461
(Singleplex)
GSK–3α⁄β  44610
(Ms)
    
GSK–3α [pY279]⁄ GSK–3β [pY216]  44604G
(Rb)
    
Hamartin  370400
(Ms)
    

 

Target ABfinity™ Recombinant Monoclonal Antibody ABfinity™ Recombinant Oligoclonal Antibody Other Primary Antibodies** Antibody Pairs ELISA Proteins Luminex® Assays
HGF    KAC2211PHG0254
PHG0321
PHG0324
LHG0071
(Singleplex)
HGH    KAQ1081PHH0014 
IGF1R 710065396700
(Ms)

See all antibodies
    
IGF1R [pYpY1135⁄1136]701067710098     
IκBα 701098710128See all antibodies KHO0211  
IκBα [pS32] 71029544725G
(Rb)
 KHO0221  
IκBα [pSpS32⁄36]  44726G
(Rb)
    
IKKα [pSpS176⁄180]  44714
(Rb)
    
IL–2701080710146AHC0722
(Ms)

See all antibodies
CRC0023KHC0021
See all ELISAs
PHC0021
PHC0023
PHC0026
PHC0027
LHC0021
(Hu Singleplex)

LMC0021
(Ms Singleplex)
IL–3  AHC0832
(Ms)

AHC0939
(Ms)
 KHC0031PHC0031
PHC0033
PHC0034
PHC0035
LHC0031
(Singleplex)
IL–6700480
701028
710042AHC0469
(Ms)

See all antibodies
CHC1263
See all antibody pairs
KHC0061
See all ELISAs
PHC0061
PHC0063
PHC0064
PHC0065
LHC0061
(Singleplex)
Insulin  180067
(Gp)
 KAQ1251 LHP0021
(Singleplex)
IR701049 AHR0221
(Ms)

See all antibodies
   LHR9111
(Singleplex)
IR [pY1158] 701070710101  KHR9121  
IR [pY1322]  44803M
(Ms)
    
IR [pY1328]  44807G
(Rb)
    
IR [pY1334]  44809G
(Rb)
 KHR9161  
IR [pY972]  44800G
(Rb)
    
IR [pYpY1162⁄1163]     KHR9131 LHR9131
(Singleplex)
IR (β–Subunit) 710076  KHR9111  
IR⁄IGF1R [pY1158]  44802G
(Rb)
    
IR⁄IGF1R [pY1162⁄pY1163]700393 44804G
(Rb)
    
IR⁄IGF1R [pYpYpY1158⁄1162⁄1163]  44806G
(Rb)
    
KIAA1303 (Raptor)  424000
(Rb)
    
LH  180037
(Ms)
    
MDM2700555710044337100
(Ms)
    
MDM2 [pS166]  441400G
(Rb)
    
mTOR  AHO1212
(Rb)

AHO1232
(Ms)
    
mTOR [pS2448] 710216441125G
(Rb)
    
NFAT1 [pS54]  44944G
(Rb)
    
NF–κB (p50)  513500
(Rb)
    
NF–κB (p65)701079710048510500
(Rb)

339900
(Ms)
 KHO0371  
NF–κB [pS529]  44711G
(Rb)
    
NF–κB [pS536]  44712G
(Rb)
    
NF–κB IA  397700
(Ms)
    
**Gp=guinea pig monoclonal IgG antibody; Ms=mouse monoclonal IgG antibody; Rb=rabbit polyclonal IgG antibody.

Find Akt Protein Targets O–V

Target ABfinity™ Recombinant Monoclonal Antibody ABfinity™ Recombinant Oligoclonal Antibody Other Primary Antibodies** Antibody Pairs ELISA Proteins Luminex® Assays
Oligomer Antibody (A11)  AHB0052
(Rb)
    
p185⁄HER–2  AHO0918
(Ms)

AHO0919
(Ms)
    
p21 701151711000
710182
337000
(Ms)
    
p21WAF1  AHZ0422
(Ms)
    
p27 719600332800
(Ms)
    
p27Kip1 [pS10]  346300
(Rb)
    
p27Kip1 [pT187] 717700379700
(Ms)
    
p27Kip1  AHZ0452
(Ms)
    
p53 [pS15]700439      
p53 [pS392]  44640G
(Rb)
    

 

Target ABfinity™ Recombinant Monoclonal Antibody ABfinity™ Recombinant Oligoclonal Antibody Other Primary Antibodies** Antibody Pairs ELISA Proteins Luminex® Assays
p53 [Thr81]  A13988
(Rb)
    
p53   491031
(Rb)

AHO0032
(Ms)

See all antibodies
    
p70S6K [pT229]  44918G
(Rb)
    
p70S6K [pT389]701064710095     
p70S6K [pTpS421⁄424] 701083710108    LHO0191
(Singleplex)
p70S6K       LHO0181
(Singleplex)
PDGFα [pY849]⁄β [pY857]  441001G
(Rb)
  PHG0035 
PDGF–D  402100
(Rb)
    
PPAP2B  391000
(Ms)
    
PRAS40 [pT246] 701058710089441100A1
(Rb)
 KHO0421  
PRAS40701075 AHO1031
(Ms)
 KHO0411 LHO0411
(Singleplex)
PTEN [pS370]  441060G
(Rb)
    
PTEN [pS385]  441064G
(Rb)
    
PTEN [pSpTpS380⁄382⁄385]  441066G
(Rb)
    
PTEN [pT383]  441062G
(Rb)
    
PTEN  512400
(Rb)

325800
(Ms)
    
Ribosomal Protein S6 [pS236]  44921G
(Rb)
    
Ribosomal Protein S6 [pSpS235⁄236]  44922G
(Rb)
    
Ribosomal Protein S6 [pSpS244⁄247]  44923G
(Rb)
    
TGF–β1   CHC1683KAC1688PHG9202
PHG9203
PHG9204
PHG9211
PHG9214
LHG0121
(Singleplex)
Tie2  425100
(Rb)
    
TIMP–1    KHC1491PHC8021
PHC8023
PHC8024
 
TrkA [pY751]  441342G
(Rb)
    
TrkA  136500
(Ms)
    
TSC2  730014
(Ms-Recombinant)

AHO1422
(Ms)
    
Tuberin  370500
(Ms)
    
VEGF Receptor–1 (Soluble)  361100
(Rb)
    
VEGF Receptor–2 [pY1054]  441046
(Rb)
    
VEGF Receptor–2 [pY1214]  441052
(Rb)
    
VEGF Receptor–2 [pYpY1054⁄1059]  441047G
(Rb)
    
VEGF Receptor–2  441053G
(Rb)

392400
(Ms)
    
VEGF Receptor–3 (C–term)  360200
(Rb)
    
VEGF–A  187328
(Ms)
    
VEGF–C (Mature)  345500
(Rb)
    
VEGF–C Propeptide  344300
(Rb)
    
VEGF–C  182255
(Rb)
    
VEGF701124710151AHG9119
(Rb)

AHG0114
(Ms)

See all antibodies
CHG0113KHG0111
See all ELISAs
PHC9391
PHC9393
PHC9394
LHG0111
(Hu Singleplex)

LMG0111
(Ms Singleplex)
VPS34 710194382100
(Rb)
    
**Gp=guinea pig monoclonal IgG antibody; Ms=mouse monoclonal IgG antibody; Rb=rabbit polyclonal IgG antibody.

Novex® Multiplex Panels for Akt Proteins

Panel Name Number of Targets Targets Polystyrene Magnetic
Human Akt Pathway-Total 7Akt, GSK-3β, IGF-1R, IR, IRS-1, PRAS40, p70-S6KLHO0002LHO0002M
Human Akt Pathway-Phospho 7Akt, GSK-3β, IGF-1R, IR, IRS-1, PRAS40, p70-S6KLHO0001LHO0001M
Human Cytokine 10-Plex 10GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNF-αLHC0001LHC0001M
Human Cytokine 25-Plex 25Eotaxin, GM-CSF, IFN-α, IFN-γ, IL-1β, IL-1RA, IL-2, IL-2R, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-15, IL-17, IP-10, MCP-1, MIG, MIP-1α, MIP-1β, RANTES, TNF-αLHC0009LHC0009M
Human Cytokine 30-Plex 30EGF, Eotaxin, FGF-Basic, G–CSF, GM–CSF, HGF, IFN-α, IFN-γ, IL–1β, IL–1RA, IL–2, IL–2R, IL–4, IL–5, IL–6, IL–7, IL–8, IL–10,  IL–12 (p40⁄ p70) IL–13, IL–15, IL–17, IP-10, MCP–1, MIG, MIP–1α, MIP–1β, RANTES, TNF-α, VEGFLHC6003LHC6003M
Human Inflammatory Cytokine 5-Plex 5GM-CSF, IL-1β, IL-6, IL-8, TNF-αLHC0003LHC0003M
Human Growth Factor 4EGF, FGF-basic, G-CSF, VEGFLHC0004 
Mouse Cytokine 10-Plex 10GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, TNF-αLMC0001LMC0001M
Mouse Cytokine 20-Plex 20FGF-basic, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, IP-10, KC, MCP-1, MIG, MIP-1α, TNF-α, VEGFLMC0006LMC0006M
Mouse Inflammatory Cytokine 4-Plex 4GM-CSF, IL-1β, IL-6, TNF-αLMC0003LMC0003M
Mouse Th1/Th2 6-Plex 6IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-12LMC0002 

* This promotion is open to customers in the US and Canada. Discount will apply to orders received by Life Technologies no later than July 31, 2013, or until promotional supplies are depleted, whichever comes first. Customer can use the discount only once. Discount only applies to a maximum of 1 kit per customer. Discount applies to list price in effect at the time order is received by Life Technologies. Cannot be combined with other discounts or promotions. Offer void where prohibited, licensed, or restricted by federal, state, provincial, or local laws or regulation or agency/institutional policy. Other restrictions may apply.

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