BioPath Online

Share |

Pathway Focus: The Akt Pathway

Antibodies for Akt Signaling Pathways ABfinity™ Technology
Antibodies for Akt Signaling Pathways—ABfinity™ Technology
Detect and Quantify Akt Pathway Markers Multiplex Luminex® AssaysDetect and Quantify Akt Pathway Markers—Multiplex Luminex® Assays

Measure Akt [pS473] and Total Protein—phosphoELISA™ Assays
Measure Akt [pS473] and Total Protein—phosphoELISA™ Assays

Visualizing Autophagy in Context of the Akt Pathway CellLight® reagents and Premo™ sensors
Visualizing Autophagy in the Context of the Akt Pathway—CellLight® reagents and Premo™ sensors
 

Cell Staining Simulation tool

Primary Antibody Search Tool

3D Cell and Cell Staining Simulation tool iPad app

Antibodies for Akt Signaling Pathways—ABfinity™ Technology

  • Lot-to-lot consistency
  • Reliable sensitivity and specificity
  • Extensive validation and characterization



ABfinity™ Recombinant Rabbit Monoclonal Antibodies represent the next generation of antibody detection tools. Produced by expressing specific cloned antibody genes in a mammalian expression system, ABfinity™ antibodies offer improved specificity and more reproducible results.

The PI3K/Akt pathway is associated with major cell functions such as cell growth, survival, apoptosis, and angiogenesis.
ABfinity™ antibodies targeting the PI3K/Akt pathway provide a critical tool for researchers investigating various diseases, including cancer, diabetes, cardiovascular conditions, and neurological disorders.

ABfinity™ antibodies have been tested for reactivity in multiple species and in multiple applications, so researchers can have greater confidence in the results. And each lot of our phosphospecific antibodies is tested in peptide competition experiments to help ensure that the antibody recognizes only the phosphorylated form of the protein.

Flow cytometry of Jurkat cells labeled with rabbit anti-AKT [pS473]
Flow cytometry of Jurkat cells labeled with rabbit anti-AKT [pS473].

Jurkat cells were incubated with (red trace) or without (green trace) 50 µM LY294002 for 1 hr prior to being fixed and permeabilized using FIX & PERM® reagents. Cells were then stained with 1 μg AKT [pS473] followed by Alexa Fluor® 488 goat anti–rabbit IgG. The blue trace represents secondary antibody alone.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
ABfinity™ Recombinant Rabbit Monoclonal Antibody Selection Guide

Description
Reactivity: Tested  (Predicted)
Applications
Quantity
Cat. No.
PKC-θ [pT538] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (X, Rt, Ms, Cp, B)
WB, F, IHC, IF/ICC
100 µg
700043
Mnk1 [pT197/pT202] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (Z, X, Sw, Rt, P, Ms, Mk (Rh), Eq, Eq, Cp, Ch, Cn, B)
WB, F, IF/ICC
100 µg
700242
Cul-2 ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu, Rt, Ms (X, Or, Mk (Rh),  Eq, Cp, Cn, B)
WB, F, IHC, IF/ICC
100 µg
700179
IRAK4 ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (Sw, Sh, Rt, Qu, Eq, Cn, B)
F, IF/ICC
100 µg
700026
AMPKβ1 [pS182] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (X, Rt, Or, Eq, Ch, Cn, B, Ms)
WB, F, IHC, IF/ICC
100 µg
700241
Smad1/5 [pS463/pS465] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (Z, X, Sw, Sh, Su, Rt, Ms, Mk (Rh), Eq, Cp, Ch, Cn, B)
F, IHC, IF/ICC
100 µg
700047
4E-BP1 [pT37] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (Z, Rt, Ms, B, Hu, Eq)
E, WB, F, IHC, IF/ICC
100 µg
700238
STAT4 ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu, Ms, Rt (Sw, Eq)
E, WB, F, IHC, IF/ICC
100 µg
700185
SUMO-3 ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu, Ms, Rt (Mk (Rh), Cp)
WB, F, IHC, IF/ICC
100 µg
700186
CASP3 [D175] ABfinity™ Recombinant Rabbit Monoclonal Antibody (clone 9H19L2)
Hu (X, Sw, Sh, Rt, Rabbit, P, Ms, Eq, Ha, Fe, Eq, Cp, Cn, B)
E, WB, F, IHC, IF/ICC
100 µg
700182

Detect and Quantify Akt Pathway Markers—Multiplex Luminex® Assays

  • Superior performance—enables accurate, reproducible, and sensitive quantitation of the Akt pathway proteins
  • High quality—in-house manufactured antibodies help ensure excellent specificity and sensitivity
  • Fast and easy protocols—perform and analyze your data typically in less than one day


Currently, Akt is one of the most actively studied kinases or kinase pathways in both the basic research and drug development arenas. Activated Akt plays a key role in mediating signals for cell growth, cell survival (anti-apoptotic), cell-cycle progression, differentiation, transcription, translation, and glucose metabolism.

Get accurate quantitation of 7 Akt pathway proteins with the AKT Total and Phospho 7-Plex Panel. The AKT 7-Plex Panel reports levels of protein, requires a small sample size, and can be run in ~4 hours.
Validation includes bead and protein specificity, precision (intra- and inter-assay, 10%), parallelism between recombinant and natural samples, linearity of dilution, recovery (>80%), and comparison to ELISA (>90%). Premixed Multiplex Kits include General Buffer Reagent Kit components and Antibody Bead Kit components, with reagents premixed for the number of markers in the assay.

Learn More about our Multiplex Protein Assays for Akt Pathway Research
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
Multiplex Luminex® Assays Selection Guide

Product Reactivity Markers
Cat. No.
Akt Pathway Phospho 7-Plex Panel
Hu, Ms, Rt
Akt [pS473], GSK-3β [pS9], IRS-1 [pS312], IGF-1R [pY1135/pY1136], IR [pY1162/pY1163], p70S6K [pTpS421/424], PRAS40 [pT246]
LHO0001
Akt Pathway Total 7-Plex Panel
Hu, Ms, Rt
Akt, GSK-3β, IGF-1R, IR, IRS-1, p70S6K, PRAS40
LHO0002

Measure Akt [pS473] and Total Protein—phosphoELISA™ Assays

  • Measure accurate levels of site-specific phosphorylation of AKT
  • More sensitive assay compared to western blots
  • Minimal cell lysate needed to get quantitative results



AKT, also known as protein kinase B a (PKB a) or RAC PKa, was initially identified as one of the downstream targets of PI3 kinase (PI3K). Eukaryotic initiation factor 4E binding protein 1 (4E-BP1) is an important effector of mTOR signaling. Hypophosphorylated 4E-BP1 acts as a translational repressor by binding and inhibiting the eIF4E. Proline-rich AKT substrate of 40 kDa (PRAS40) is a direct substrate for AKT.

AKT can be activated by a diverse array of growth factors and physiological stimuli in a PI3K- dependent manner.
Activated AKT affects downstream markers such as 4E-BP1 and PRAS40, and acts as a key mediator of signals for cell survival, proliferation, and angiogenesis.

Invitrogen offers sandwich ELISA kits to quickly detect and quantify phosphorylation site–specific proteins in cell lysates. Total protein kits are provided to normalize the protein content of the sample, independent of the phosphorylation state. Our ELISA kits deliver quantitative results and give you workflow efficiencies with high sample throughput and a 96-well plate format. Results are achieved after as little time as 4 hours of incubation, and assays require less sample than western blots because of the improved sensitivity.

  • Learn More about Sandwich ELISA Kits for Akt Pathway Research
4E-BP1 [pT46] and 4E-BP1 (total) ELISAs on HEK 293 cells
4E-BP1 [pT46] and 4E-BP1 (total) ELISAs on HEK 293 cells.
HEK 293 cells were serum-starved overnight, then deprived of amino acids in PBS for 60 minutes. Untreated HEK 293 cells were used as control. Cell extracts were prepared and analyzed with the 4E-BP1 [pT46] and 4E-BP1 (Total) ELISA. The results show that compared to untreated control, the Thr46 phosphorylation of 4E-BP1 is down-regulated in amino acid–starved HEK 293 cells, whereas the total level of 4E-BP1 remains relatively constant.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
phosphoELISA™ Assays Selection Guide

Product
Species
Quantity
Cat. No.
Akt [pS473] ELISA Kit
Hu, Ms, Rt
96 tests
KHO0111
Akt [pS473] High Sensitivity ELISA Kit
Hu, Ms, Rt
96 testsKHO0541
Akt [pT308] ELISA Kit
Hu
96 testsKHO0201
Akt (Total) ELISA Kit
Hu, Ms, Rt
96 testsKHO0101
4E-BP1 [pT46] ELISA Kit
Hu, Ms, Rt
96 testsKHO0691
4E-BP1 (Total) ELISA Kit
Hu, Ms, Rt
96 testsKHO0681
PRAS40 [pT246] ELISA Kit
Hu
96 testsKHO0421
PRAS40 (Total) ELISA Kit
Hu
96 testsKHO0411

Visualizing Autophagy in the Context of the Akt Pathway—CellLight® Reagents and Premo™ Sensors

  • Track autophagy from start to finish in live cells—from the formation of autophagosomes to the autophagosome fusion with lysosomes
  • Visualize autophagy in physiologically relevant models, stem cells, and primary cells (including neurons)
  • Excellent lot-to-lot consistency with ABfinity™ technology

 



The BacMam 2.0–based CellLight® reagents and Premo™ sensors are fluorescent protein–signal peptide fusions that permit accurate and specific targeting to cellular structures. They use a modified insect cell baculovirus coupled with a mammalian promoter as a vehicle to efficiently deliver and express genes in mammalian cells. Unlike expression vectors, BacMam reagents provide a drastically simplified workflow, enable titratable and reproducible expression, and offer high cotransduction efficiency.

 

This enables multiple BacMam reagents to be used in the same cell (including primary and stem cells) for live-cell imaging and for fixed-cell analyses following formaldehyde-based fixation.

In a recent report, inhibition of AKT was shown not to induce significant apoptosis, but did promote autophagy (J Cell Biol 183:101–116 (2008)).

Premo™ sensors and CellLight® reagents enable researchers to accurately image autophagy via LC3B recruitment to autophagosomes, an early step in autophagy that occurs when the protein is cleaved and lipidated, as well as visualization of key organelles associated with autophagy, including (but not limited to) mitochondria, peroxisomes, and the Golgi apparatus.

Multicolor imaging of autophagosomes and other organelles
Multicolor imaging of autophagosomes and other organelles.
A549 cells were transduced with the Premo™ Autophagy Sensor LC3B-GFP and CellLight® reagents to label specific organelles. At 24 hr after transduction, cells were treated with 50 µM chloroquine overnight to favor autophagosome formation. Nuclei were labeled with Hoechst 33342 (blue). Cells were transduced with (A) CellLight® Mitochondria-RFP, (B) CellLight® Golgi-RFP, (C) CellLight® Lysosomes-GFP, and (D) CellLight® ER-RFP.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
CellLight® Reagents and Premo™ Sensors Selection Guide

Product
  Cat. No.
Premo™ Autophagy Sensor LC3B-GFP *BacMam 2.0*

P36235
Premo™ Autophagy Sensor LC3B-RFP *BacMam 2.0*

P36236
LC3B Antibody Kit for Autophagy

L10382
CellLight® Golgi-GFP *BacMam 2.0*

C10592
CellLight® Golgi-RFP *BacMam 2.0*

C10593
CellLight® Lysosomes-GFP *BacMam 1.0*

C10596
CellLight® Lysosomes-RFP *BacMam 2.0*

C10597
CellLight® Mitochondria-GFP *BacMam 2.0*

C10600
CellLight® Mitochondria-RFP *BacMam 2.0*

C10601
CellLight® Peroxisomes-GFP *BacMam 2.0*

C10604