Name: PI3K/AKT signalling pathway panel
SKU: ab283852
Rating: 5 (1 reviews)

PI3K/AKT signalling pathway panel containing 4 antibodies against PI 3 Kinase p85 alpha, AKT1, AKT2, AKT3 and mTOR

- Provided as a sampler panel to allow you to easily evaluate each in your required applications.

Images

Immunocytochemistry - PI3K/AKT signalling pathway panel (AB283852), expandable thumbnail

Publications

Key facts

Reactive species: Mouse, Human, Rat
Target: PIK3R1
(See target data below)

Target data

See full target information PIK3R1

Function

Binds to activated (phosphorylated) protein-Tyr kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Necessary for the insulin-stimulated increase in glucose uptake and glycogen synthesis in insulin-sensitive tissues. Plays an important role in signaling in response to FGFR1, FGFR2, FGFR3, FGFR4, KITLG/SCF, KIT, PDGFRA and PDGFRB. Likewise, plays a role in ITGB2 signaling (PubMed:17626883, PubMed:19805105, PubMed:7518429). Modulates the cellular response to ER stress by promoting nuclear translocation of XBP1 isoform 2 in a ER stress- and/or insulin-dependent manner during metabolic overloading in the liver and hence plays a role in glucose tolerance improvement (PubMed:20348923).

Additional Targets

MTOR, AKT1

Alternative names

GRB1, PIK3R1, Phosphatidylinositol 3-kinase regulatory subunit alpha, PI3-kinase regulatory subunit alpha, PI3K regulatory subunit alpha, PtdIns-3-kinase regulatory subunit alpha, Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha, PI3-kinase subunit p85-alpha, PtdIns-3-kinase regulatory subunit p85-alpha

What's included?

1 Kit

Components

ab192623

Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853]

2 x 10 µL

ab191606

Anti-PI 3 Kinase p85 alpha antibody [EPR18702]

2 x 10 µL

ab134903

Anti-mTOR antibody [EPR390(N)]

2 x 10 µL

ab179463

Rabbit monoclonal [EPR16798] to AKT1 + AKT2 + AKT3

2 x 10 µL

Key facts

Reactive species: Mouse, Human, Rat
Target: PIK3R1

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: -20°C
Appropriate long-term storage conditions: -20°C
Storage information: -20°C

Notes

Product Specifications
PI3K/AKT signalling pathway panel (ab283852) is a panel of 5 antibodies specific to the PI3K/AKT signalling pathway, which regulates cell frowth, survival, metabolism and proliferation.
This panel contains 4 rabbit recombinant monoclonal antibodies and 1 rabbit polyclonal antibody to PI 3 Kinase p85 alpha, PI 3 Kinase p85 alpha (pY607), [AKT1 + AKT2 + AKT3], [AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472)] and mTOR in convenient trial sizes (20 µL) to allow for efficient and cost-effect testing of the antibodies simultaneously.

The individual antibodies included in this panel are:

Rabbit recombinant monoclonal anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Rabbit polyclonal anti-PI 3 Kinase p85 alpha (phospho Y607) antibody (ab182651)
Rabbit recombinant monoclonal anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)
Rabbit recombinant monoclonal anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] (ab192623)
Rabbit recombinant monoclonal anti-mTOR antibody [EPR390(N)] (ab134903)

All of these component antibodies are available for purchase individually using their corresponding abIDs.

Quality and Validation
Abcams high quality validation processes ensure each component antibody is sensitive and specific to the target protein.
All antibodies in this panel have been validated in human, mouse and rat samples in a variety of applications, and some also have reactivity in other species.
For information on the species reactivity and application validation of each antibody within the panel, please consult the individual datasheet for each antibody. Suggested dilution factor and additional information on protocols can also be found on the antibody datasheets.
All antibodies in this panel have a minimum of 60 citations in peer-reviewed journals and are trusted by scientific community.

Related Products
Many of our antibody clones are also available as carrier-free formulations for easy conjugation to labels of your choice, or in pre-conjugated formats. Please search the clone name for alternative formulations and related products.

We recommend using ab283852 together with ab205718.

Explore our range of antibody sample panels designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format.

Directly conjugated versions of our antibodies are available and ready to use for multicolor flow cytometry or immunocytochemistry analysis.

Carrier-free formulations of our recombinant antibodies are also available for easy conjugation to labels of your choice and for multiplex applications. Use our intuitive search and select carrier-free or your label of choice. For bespoke conjugations or large volumes email bespoke@abcam.com.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MTOR also known as mechanistic target of rapamycin is a protein kinase involved in regulating cell growth proliferation motility and survival. It has a molecular weight of approximately 289 kDa and is expressed widely across various tissues notably in areas of high cellular activity. mTOR is part of the PI3K/AKT signaling pathway where it functions as a central regulator. PI3 kinase p85 alpha is a regulatory subunit of phosphoinositide 3-kinases which plays an important role in the modulation of AKT signaling. AKT1 AKT2 and AKT3 are serine/threonine-specific protein kinases that play an important role in transmitting signals downstream of PI3K with each isoform having unique tissue-specific roles.

Biological function summary

MTOR functions as a core component of two distinct complexes: mTORC1 and mTORC2. mTORC1 regulates protein synthesis by modulating the phosphorylation of key translational regulators while mTORC2 is involved in the regulation of the actin cytoskeleton. The PI3 kinase p85 alpha a regulatory entity mediates signaling through its interaction with catalytic subunits impacting glucose metabolism and growth. AKT being a pivotal node in cellular processes modulates glucose uptake and cell proliferation and prevents apoptosis through its activity. Collectively these proteins act to integrate signals from growth factors and nutrients to enact biological responses critical for cellular homeostasis.

Pathways

MTOR PI3 kinase p85 alpha and AKT isoforms are important components of the PI3K/AKT/mTOR pathway. This pathway plays a significant role in cellular metabolism growth survival and angiogenesis. Within this context mTOR acts downstream of AKT to promote protein synthesis and cell growth. PI3K signaling initiates AKT activation which then phosphorylates various substrates including mTOR. Members of this pathway such as PTEN regulate the pathway's activity negatively modulating cellular responses and ensuring proper cell function and survival.

Associated diseases and disorders

The dysregulation of mTOR PI3K and AKT is associated with cancer and type 2 diabetes. These proteins when aberrantly activated can lead to uncontrolled cell proliferation and tumorigenesis with mTOR being a target for rapamycin an anti-cancer drug. In type 2 diabetes insulin signaling involving the PI3K/AKT pathway becomes impaired leading to improper glucose homeostasis. The intricate relations of these proteins within these diseases highlight their potential as targets for therapeutic interventions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

Immunocytochemistry - PI3K/AKT signalling pathway panel (ab283852)
Immunocytochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells, untreated or treated with PDGF (100 ng/ml) for 1 hour, labeling AKT3 (phospho S472) + AKT2 (phospho S474) + AKT1 (phospho S473) with Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] ab192623 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
The signal increased after treatment with PDGF (100 ng/ml) for 1 hour on NIH/3T3 cells.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Immunoprecipitation - PI3K/AKT signalling pathway panel (ab283852)
KT3 (phospho S472) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] ab192623 at 1/40 dilution. Western blot was performed from the immunoprecipitate using Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] ab192623 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10μg (Input).
Lane 2: Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] ab192623 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] ab192623 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
Western blot - PI3K/AKT signalling pathway panel (ab283852)
All lanes: Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] ab192623) at 1/1000 dilution.
Lane 1: LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates, 20 μg
Lane 2: LNCaP (Human prostate carcinoma epithelial cell) treated with 0.1 uM Calyculin A for 30 minutes whole cell lysates, 20 μg
Lane 3: A549 (Human lung carcinoma epithelial cell) whole cell lysates, 20 μg
Lane4: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates, 20 μg
Lane 5: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates, 20 μg
Lane6: HUVEC (Human umbilical vein endothelial cell) whole cell lysates, 20 μg
Lane7: C2C12 (Mouse myoblasts myoblast) whole cell lysates, 20 μg
Lane 8: Mouse brain lysates, 20 μg
Lane9: Rat heart lysates, 20 μg
Secondary (all lanes): Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted MW: 56 kDa.
Observed MW: 56 kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 50 seconds.
Western blot - PI3K/AKT signalling pathway panel (ab283852)
Lanes 1 - 3: Merged signal (red and green). Green - Anti-mTOR antibody [EPR390(N)] ab134903 observed at 289 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.
Lane 1: Wild-type HEK293T whole cell lysate, 20 μg
Lane 2: MTOR knockout HEK293T whole cell lysate, 20 μg
Lane 3: K562 whole cell lysate, 20 μg
Anti-mTOR antibody [EPR390(N)] ab134903 was shown to specifically react with mTOR in wild-type HEK293T cells as signal was lost in MTOR knockout cells. Wild-type and MTOR knockout samples were subjected to SDS-PAGE. Anti-mTOR antibody [EPR390(N)] ab134903 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry - PI3K/AKT signalling pathway panel (ab283852)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling PI3K p85 with Anti-PI 3 Kinase p85 alpha antibody [EPR18702] ab191606 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor^®594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: Anti-PI 3 Kinase p85 alpha antibody [EPR18702] ab191606 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
Immunocytochemistry - PI3K/AKT signalling pathway panel (ab283852)
Immunocytochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling AKT1 + AKT2 + AKT3 with Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab179463 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on K562 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
1. Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab179463 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.
Western blot - PI3K/AKT signalling pathway panel (ab283852)
Lanes 1 - 3: Merged signal (red and green). Green - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] ab191606 observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
Lane 1: Wild type HAP1 whole cell lysate, 20 μg
Lane 2: PIK3R1 knockout HAP1 whole cell lysate, 20 μg
Lane 3: Jurkat whole cell lysate, 20 μg
Anti-PI 3 Kinase p85 alpha antibody [EPR18702] ab191606 was shown to specifically react with PIK3R1 when PIK3R1 knockout samples were used. Wild-type and PIK3R1 knockout samples were subjected to SDS-PAGE. Anti-PI 3 Kinase p85 alpha antibody [EPR18702] ab191606 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - PI3K/AKT signalling pathway panel (ab283852)
All lanes: Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab179463) at 1/1000 dilution.
Lane 1: MCF7 (Human breast adenocarcinoma cell line) whole cell lysates, 20 μg
Lane 2: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates, 20 μg
Lane 3: Hep G2 (Human liver hepatocellular carcinoma) whole cell lysates, 20 μg
Secondary (all lanes): Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution.
Predicted MW: 56 kDa.
Observed MW: 56 kDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 70 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PI3K/AKT signalling pathway panel (ab283852)
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling AKT1 + AKT2 + AKT3 with Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Human renal cortex is observed. Counterstained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PI3K/AKT signalling pathway panel (ab283852)
Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling AKT1 + AKT2 + AKT3 with Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Rat cerebral cortex is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - PI3K/AKT signalling pathway panel (ab283852)
Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PI3K p85 with Anti-PI 3 Kinase p85 alpha antibody [EPR18702] ab191606 at 1/150 dilution (red) compared with a Rabbit IgG,monoclonal - Isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - PI3K/AKT signalling pathway panel (ab283852)
Flow cytometric analysis of 2% paraformaldehyde-fixed A549 (Human lung carcinoma) cells labeling AKT1 + AKT2 + AKT3 with Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab179463 at 1/330 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.