Akt Signaling Pathway—Novex® Antibodies, ELISAs, & Luminex® Assays

Akt Pathway Overview

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

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Introductory Trial Offer

The Best Way to Put Your Pathway Research to the Test

For a limited time we are offering an introductory trial offer, the best way to evaluate tools for your AKT/mTOR research, to keep your research a step ahead. Select any Novex® AKT/mTOR pathway–related product on the page, and save 50%. Use promotion code P923057 (US) or P923063 (Canada) when placing your order.*

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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).

Western Blot Analysis

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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

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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

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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	Luminex® Assays
4E–BP1			AHO1382 (Ms) See all antibodies		KHO0681
4E–BP1 [pT37]	700238
4E–BP1 [pT46]	700397		441170G (Rb)		KHO0691
Akt		710005	44609M (Ms) See all antibodies		KHO0101 KHO0531 See all ELISAs	LHO0091 (Singleplex)
Akt [pS473] & [npS473]	700392 700256		44622M [npS473] (Ms) 44621G (Rb)	CHO0115	KHO0111 KHO0541 See all ELISAs	LHO0101 (Singleplex)
Akt [pT308]	701052	710122	44602G (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–Met	700261	718000	182257 (Rb) 370100 (Ms) See all antibodies	CHO0285	KHO0251
c–Met (soluble)				CHO0315	KHO2031
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		712700	AHO0462 (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)
EGFR	700308 700994		44799G (Rb) AHR5062 (Ms) See all antibodies		KHR9061 See all ELISAs		LHR9061 (Singleplex)
EGFR [pY845]			44784G (Rb)
EGFR [pY992]			44786G (Rb)
EGFR [pY1068]	701053	710123	44788G (Rb)		KHR9081		LHR9081 (Singleplex)
EGFR [pY1086]			44790G (Rb) 369700 (Rb)
EGFR [pY1148]		710213	44792G (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.

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Find Akt Protein Targets F–N

Target	ABfinity™ Recombinant Monoclonal Antibody	ABfinity™ Recombinant Oligoclonal Antibody	Other Primary Antibodies**	ELISA	Proteins	Luminex® Assays
FGF–basic		710148		KHG0021 (96 Tests) KHG0022 (192 Tests)	13256029	LHG0021 (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–3β	701102	710132	399500 (Ms) See all antibodies	KHO0451		LHO0451 (Singleplex)
GSK–3β [pS9]	701069	710100		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					KAC2211	PHG0254 PHG0321 PHG0324	LHG0071 (Singleplex)
HGH					KAQ1081	PHH0014
IGF1R		710065	396700 (Ms) See all antibodies
IGF1R [pYpY1135⁄1136]	701067	710098
IκBα	701098	710128	See all antibodies		KHO0211
IκBα [pS32]		710295	44725G (Rb)		KHO0221
IκBα [pSpS32⁄36]			44726G (Rb)
IKKα [pSpS176⁄180]			44714 (Rb)
IL–2	701080	710146	AHC0722 (Ms) See all antibodies	CRC0023	KHC0021 See all ELISAs	PHC0021 PHC0023 PHC0026 PHC0027	LHC0021 (Hu Singleplex) LMC0021 (Ms Singleplex)
IL–3			AHC0832 (Ms) AHC0939 (Ms)		KHC0031	PHC0031 PHC0033 PHC0034 PHC0035	LHC0031 (Singleplex)
IL–6	700480 701028	710042	AHC0469 (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)
IR	701049		AHR0221 (Ms) See all antibodies				LHR9111 (Singleplex)
IR [pY1158]	701070	710101			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)
MDM2	700555	710044	337100 (Ms)
MDM2 [pS166]			441400G (Rb)
mTOR			AHO1212 (Rb) AHO1232 (Ms)
mTOR [pS2448]		710216	441125G (Rb)
NFAT1 [pS54]			44944G (Rb)
NF–κB (p50)			513500 (Rb)
NF–κB (p65)	701079	710048	510500 (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.

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Find Akt Protein Targets O–V

Target	ABfinity™ Recombinant Monoclonal Antibody	ABfinity™ Recombinant Oligoclonal Antibody	Other Primary Antibodies**
Oligomer Antibody (A11)			AHB0052 (Rb)
p185⁄HER–2			AHO0918 (Ms) AHO0919 (Ms)
p21	701151	711000 710182	337000 (Ms)
p21WAF1			AHZ0422 (Ms)
p27		719600	332800 (Ms)
p27Kip1 [pS10]			346300 (Rb)
p27Kip1 [pT187]		717700	379700 (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]	701064	710095
p70S6K [pTpS421⁄424]	701083	710108					LHO0191 (Singleplex)
p70S6K							LHO0181 (Singleplex)
PDGFα [pY849]⁄β [pY857]			441001G (Rb)			PHG0035
PDGF–D			402100 (Rb)
PPAP2B			391000 (Ms)
PRAS40 [pT246]	701058	710089	441100A1 (Rb)		KHO0421
PRAS40	701075		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				CHC1683	KAC1688	PHG9202 PHG9203 PHG9204 PHG9211 PHG9214	LHG0121 (Singleplex)
Tie2			425100 (Rb)
TIMP–1					KHC1491	PHC8021 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)
VEGF	701124	710151	AHG9119 (Rb) AHG0114 (Ms) See all antibodies	CHG0113	KHG0111 See all ELISAs	PHC9391 PHC9393 PHC9394	LHG0111 (Hu Singleplex) LMG0111 (Ms Singleplex)
VPS34		710194	382100 (Rb)
**Gp=guinea pig monoclonal IgG antibody; Ms=mouse monoclonal IgG antibody; Rb=rabbit polyclonal IgG antibody.

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Novex® Multiplex Panels for Akt Proteins

Panel Name	Number of Targets	Targets	Polystyrene	Magnetic
Human Akt Pathway-Total	7	Akt, GSK-3β, IGF-1R, IR, IRS-1, PRAS40, p70-S6K	LHO0002	LHO0002M
Human Akt Pathway-Phospho	7	Akt, GSK-3β, IGF-1R, IR, IRS-1, PRAS40, p70-S6K	LHO0001	LHO0001M
Human Cytokine 10-Plex	10	GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNF-α	LHC0001	LHC0001M
Human Cytokine 25-Plex	25	Eotaxin, 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-α	LHC0009	LHC0009M
Human Cytokine 30-Plex	30	EGF, 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, VEGF	LHC6003	LHC6003M
Human Inflammatory Cytokine 5-Plex	5	GM-CSF, IL-1β, IL-6, IL-8, TNF-α	LHC0003	LHC0003M
Human Growth Factor	4	EGF, FGF-basic, G-CSF, VEGF	LHC0004
Mouse Cytokine 10-Plex	10	GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, TNF-α	LMC0001	LMC0001M
Mouse Cytokine 20-Plex	20	FGF-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, VEGF	LMC0006	LMC0006M
Mouse Inflammatory Cytokine 4-Plex	4	GM-CSF, IL-1β, IL-6, TNF-α	LMC0003	LMC0003M
Mouse Th1/Th2 6-Plex	6	IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-12	LMC0002

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* 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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