AKT Serine/Threonine Kinase 1 Peptide 5


Background

Catalog Number:

CPTC-AKT1-4

RRID:

AB_2827858

Target Antigen:

AKT Serine/Threonine Kinase 1 Peptide 5

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

11/21/2023

Antigen Recognition(s):

Peptide, Recombinant Full-length, Endogenous, Phosphorylation

Thematic Panel(s):

Ras Pathway

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Publications
Characterization Data [Compare Characterization Data]
  IHC Tissue
Click to enlarge image Tissue Micro-Array (TMA) core of colon cancer showing cytoplasmic staining using Antibody CPTC-AKT1-4. Titer: 1:1000 Click image to enlarge

CPTC-AKT1-4 IHC Tissue

Result: Presumed Positive

Tissue Micro-Array (TMA) core of colon cancer showing cytoplasmic staining using Antibody CPTC-AKT1-4. Titer: 1:1000


  IP
Click to enlarge image Immuno-precipitation performed using CPTC-AKT1-4 as capture antibody against MCF7 and HeLa lysates.  Eluates were tested in automated western blot using the same CPTC-AKT1-4 as detection antibody. The target protein was detected only in the whole lysates (lanes MCF7 SM, HeLa SM), and in the IP eluates (lane MCF7 – IP – AKT1-4, HeLa – IP – AKT1-4). Click image to enlarge

CPTC-AKT1-4 – IP-Wes

Result: Positive

Immuno-precipitation performed using CPTC-AKT1-4 as capture antibody against MCF7 and HeLa lysates. Eluates were tested in automated western blot using the same CPTC-AKT1-4 as detection antibody. The target protein was detected only in the whole lysates (lanes MCF7 SM, HeLa SM), and in the IP eluates (lane MCF7 – IP – AKT1-4, HeLa – IP – AKT1-4).


  Immuno-MRM
Click to enlarge image iMRM screening results for clone CPTC-AKT1-4. The clone is able to selectively pull down the target peptide (CPTC-AKT1 Peptide 5, (pT)FCGTPEYLAPEVLEDNDYGRK)

Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html) Click image to enlarge

CPTC-AKT1-4 iMRM (synthetic peptide)

Result: Positive

iMRM screening results for clone CPTC-AKT1-4. The clone is able to selectively pull down the target peptide (CPTC-AKT1 Peptide 5, (pT)FCGTPEYLAPEVLEDNDYGRK)

Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html)


  Immunofluorescence

CPTC-AKT1-4 Immunofluorescence

Result: Negative

Immunofluorescence staining of MCF10a cells stably transfected with KRAS (MCF10a-KRAS) using CPTC-AKT1-4 antibody. AKT1 protein expression was not detected.


  Immunofluorescence

CPTC-AKT1-4 Immunofluorescence

Result: Negative

Immunofluorescence staining using CPTC-AKT1-4 as primary antibody against MCF7 cells shows no localization of AKT1 protein.


  NCI 60 Protein Array

CPTC-AKT1-4 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western Blot using CPTC-AKT1-4 as primary Ab against recombinant AKT1 (lane 2). Also included are molecular wt. standards (lane 1). Click image to enlarge

CPTC-AKT1-4 Western Blot

Result: Positive

Western Blot using CPTC-AKT1-4 as primary Ab against recombinant AKT1 (lane 2). Also included are molecular wt. standards (lane 1).


  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Automated western blot using CPTC-AKT1-4 as primary antibody against recombinant AKT1. The antibody can recognize the target. Click image to enlarge

CPTC-AKT1-4 automated western blot (recombinant protein)

Result: Positive

Automated western blot using CPTC-AKT1-4 as primary antibody against recombinant AKT1. The antibody can recognize the target.


  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western blot using CPTC-AKT1-4 as primary antibody against recombinant AKT1. The antibody can recognize the target. Click image to enlarge

CPTC-AKT1-4 western blot (recombinant protein)

Result: Positive

Western blot using CPTC-AKT1-4 as primary antibody against recombinant AKT1. The antibody can recognize the target.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western Blot using CPTC-AKT1-1 as primary Ab against MCF10A-KRas cell lysate (lane 2). Also included are molecular wt. standards Click image to enlarge

CPTC-AKT1-4 Cell Lysate Western Blot

Result: Positive

Western Blot using CPTC-AKT1-1 as primary Ab against MCF10A-KRas cell lysate (lane 2). Also included are molecular wt. standards


  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated western blot using CPTC-AKT1-4 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody cannot recognize the target in the cell lysates. Click image to enlarge

CPTC-AKT1-4 automated western blot (lysates)

Result: Negative

Automated western blot using CPTC-AKT1-4 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody cannot recognize the target in the cell lysates.


  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-AKT1-4 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody presumably recognizes the target in the MDA-MB-231, MCF7, T47D and HeLa cell lysates. Click image to enlarge

CPTC-AKT1-4 western blot (lysates)

Result: Presumed Positive (with additional bands)

Western blot using CPTC-AKT1-4 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody presumably recognizes the target in the MDA-MB-231, MCF7, T47D and HeLa cell lysates.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Single cell western blot using CPTC-AKT1-4 as a primary antibody against cell lysates.  Phosphorylated AKT1 protein was detected.  Relative expression of total AKT1 in MCF7 cells (A).  Percentage of cells that express AKT1 (B).  Average expression of AKT1 protein per cell (C).  All data is normalized to β-tubulin expression. Click image to enlarge

CPTC-AKT1-4 Single Cell Western Blot (Cell Lysate)

Result: Positive

Single cell western blot using CPTC-AKT1-4 as a primary antibody against cell lysates. Phosphorylated AKT1 protein was detected. Relative expression of total AKT1 in MCF7 cells (A). Percentage of cells that express AKT1 (B). Average expression of AKT1 protein per cell (C). All data is normalized to β-tubulin expression.


Background

Catalog Number:

CPTC-AKT1-5

RRID:

AB_2827859

Target Antigen:

AKT Serine/Threonine Kinase 1 Peptide 5

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

07/14/2022

Antigen Recognition(s):

Peptide, Recombinant Full-length, Phosphorylation

Thematic Panel(s):

Ras Pathway

Purchase
Publications
Characterization Data [Compare Characterization Data]
  Immuno-MRM
Click to enlarge image iMRM screening results for clone CPTC-AKT1-5. The clone is able to pull down the target peptide (panel 1, CPTC-AKT1 Peptide 5, (pT)FCGTPEYLAPEVLEDNDYGRK) and the correspondent non-phosphorylated peptide (panel 2, TFCGTPEYLAPEVLEDNDYGR)

Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html) Click image to enlarge

CPTC-AKT1-5 iMRM (synthetic peptide)

Result: Positive

iMRM screening results for clone CPTC-AKT1-5. The clone is able to pull down the target peptide (panel 1, CPTC-AKT1 Peptide 5, (pT)FCGTPEYLAPEVLEDNDYGRK) and the correspondent non-phosphorylated peptide (panel 2, TFCGTPEYLAPEVLEDNDYGR)

Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html)


  NCI 60 Protein Array

CPTC-AKT1-5 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Automated western blot using CPTC-AKT1-5 as primary antibody against recombinant AKT1. The antibody can recognize the target. Click image to enlarge

CPTC-AKT1-5 automated western blot (recombinant protein)

Result: Positive

Automated western blot using CPTC-AKT1-5 as primary antibody against recombinant AKT1. The antibody can recognize the target.


  Western Blot - Recombinant Protein or Peptide

CPTC-AKT1-5 western blot (recombinant protein)

Result: Negative

Western blot using CPTC-AKT1-5 as primary antibody against recombinant AKT1. The antibody cannot recognize the target.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated western blot using CPTC-AKT1-5 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody cannot recognize the target in the cell lysates. Click image to enlarge

CPTC-AKT1-5 automated western blot (lysates)

Result: Negative

Automated western blot using CPTC-AKT1-5 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody cannot recognize the target in the cell lysates.


  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-AKT1-5 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody cannot recognize the target in the cell lysates. Click image to enlarge

CPTC-AKT1-5 western blot (lysates)

Result: Negative

Western blot using CPTC-AKT1-5 as primary antibody against whole lysates of cell lines cell MDA-MB-231, HT-29, MCF7, T47D, SK-OV-3, and HeLa. The antibody cannot recognize the target in the cell lysates.


Characterization SOP Files

Background

NCI Identification Number:

00227

Antigen Name:

AKT Serine/Threonine Kinase 1 Peptide 5

CPTC Name:

CPTC-AKT1 Peptide 5

Aliases:

AKT Serine/Threonine Kinase 1; V-Akt Murine Thymoma Viral Oncogene Homolog 1; Protein Kinase B Alpha; Proto-Oncogene C-Akt; RAC-PK-Alpha; EC 2.7.11.1; PKB Alpha; PKB; RAC; V-Akt Murine Thymoma Viral Oncogene-Like Protein 1; RAC-Alpha Serine/Threonine-Protein Kinase; Serine-Threonine Protein Kinase; Rac Protein Kinase Alpha; Protein Kinase B; PKB-ALPHA; RAC-ALPHA; EC 2.7.11; AKT1m; PRKBA; CWS6; AKT

Function:

The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Mutations in this gene have been associated with the Proteus syndrome. Multiple alternatively spliced transcript variants have been found for this gene.

AKT1 (AKT Serine/Threonine Kinase 1) is a Protein Coding gene. Diseases associated with AKT1 include Proteus Syndrome, Somatic and Cowden Syndrome 6. Among its related pathways are PI-3K cascade-FGFR3 and GDNF-Family Ligands and Receptor Interactions. GO annotations related to this gene include identical protein binding and protein kinase activity. An important paralog of this gene is AKT3.

AKT1 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT is responsible of the regulation of glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface. Phosphorylation of PTPN1 at Ser-50 negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling. Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport. AKT regulates also the storage of glucose in the form of glycogen by phosphorylating GSK3A at Ser-21 and GSK3B at Ser-9, resulting in inhibition of its kinase activity. Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven. AKT regulates also cell survival via the phosphorylation of MAP3K5 (apoptosis signal-related kinase). Phosphorylation of Ser-83 decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at Ser-939 and Thr-1462, thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization. In particular, FOXO1 is phosphorylated at Thr-24, Ser-256 and Ser-319. FOXO3 and FOXO4 are phosphorylated on equivalent sites. AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1. AKT phosphorylates Ser-454 on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis. Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of Ser-273, resulting in reduced cyclic AMP levels and inhibition of lipolysis. Phosphorylates PIKFYVE on Ser-318, which results in increased PI(3)P-5 activity. The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth. AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation. Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). AKT mediates the antiapoptotic effects of IGF-I. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. May be involved in the regulation of the placental development. Phosphorylates STK4/MST1 at Thr-120 and Thr-387 leading to inhibition of its: kinase activity, nuclear translocation, autophosphorylation and ability to phosphorylate FOXO3. Phosphorylates STK3/MST2 at Thr-117 and Thr-384 leading to inhibition of its: cleavage, kinase activity, autophosphorylation at Thr-180, binding to RASSF1 and nuclear translocation. Phosphorylates SRPK2 and enhances its kinase activity towards SRSF2 and ACIN1 and promotes its nuclear translocation. Phosphorylates RAF1 at Ser-259 and negatively regulates its activity. Phosphorylation of BAD stimulates its pro-apoptotic activity. Phosphorylates KAT6A at Thr-369 and this phosphorylation inhibits the interaction of KAT6A with PML and negatively regulates its acetylation activity towards p53/TP53.
AKT1-specific substrates have been recently identified, including palladin (PALLD), which phosphorylation modulates cytoskeletal organization and cell motility; prohibitin (PHB), playing an important role in cell metabolism and proliferation; and CDKN1A, for which phosphorylation at Thr-145 induces its release from CDK2 and cytoplasmic relocalization. These recent findings indicate that the AKT1 isoform has a more specific role in cell motility and proliferation. Phosphorylates CLK2 thereby controlling cell survival to ionizing radiation.

Akt (Protein kinase B, PKB) is a serine/threonine kinase that plays a key in regulating cell survival, insulin signaling, angiogenesis and tumor formation. Akt is a downstream mediator of the PI 3-K pathway, which results in the recruitment of Akt to the plasma membrane.

Chromosomal Localization:

14q32.33

Accession Number:

NP_001014431.1

UniProt Accession Number:

P31749

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

(pT)FCGTPEYLAPEVLEDNDYGR

Native Sequence:

Calculated Isoelectric Point:

0

Molecular Weight:

2310

Last Updated:

10/01/2019

Links

Characterization Data

SOPs

No SOPs available.

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