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Phospho-AKT1 (Ser473) Recombinant Monoclonal Antibody

Datasheet
Code CSB-RA001553A473phHU
Size US$210
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Image
  • Western Blot
    Positive WB detected in 293 whole cell lysate(treated with Calyculin A or not)
    All lanes Phospho-AKT1 antibody at 1.08μg/ml
    Secondary
    Goat polyclonal to rabbit IgG at 1/50000 dilution
    Predicted band size: 60 KDa
    Observed band size: 60 KDa
  • IHC image of CSB-RA001553A473phHU diluted at 1:100 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
  • IHC image of CSB-RA001553A473phHU diluted at 1:100 and staining in paraffin-embedded human lung cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
  • Immunofluorescence staining of Hela cells(treated with 100mM EGF for 20min) with CSB-RA001553A473phHU at 1:68,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4℃. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG (H+L).
  • Immunoprecipitating Phospho-AKT1 in 293 whole cell lysate treated with Calyculin A
    Lane 1: Rabbit control IgG(1μg)instead of CSB-RA001553A473phHU in 293 whole cell lysate treated with Calyculin A. For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
    Lane 2: CSB-RA001553A473phHU(3μg)+ 293 whole cell lysate treated with Calyculin A(1mg)
    Lane 3: 293 whole cell lysate treated with Calyculin A (20μg)
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Product Details

Uniprot No.
P31749
Target Names
AKT1
Alternative Names
AKT 1 antibody; AKT antibody; AKT1 antibody; AKT1_HUMAN antibody; C AKT antibody; cAKT antibody; MGC99656 antibody; PKB alpha antibody; PKB antibody; PKB-ALPHA antibody; PRKBA antibody; Protein Kinase B Alpha antibody; Protein kinase B antibody; Proto-oncogene c-Akt antibody; RAC Alpha antibody; RAC antibody; Rac protein kinase alpha antibody; RAC Serine/Threonine Protein Kinase antibody; RAC-alpha serine/threonine-protein kinase antibody; RAC-PK-alpha antibody; v akt murine thymoma viral oncogene homolog 1 antibody; vAKT Murine Thymoma Viral Oncogene Homolog 1 antibody
Species Reactivity
Human
Immunogen
A synthesized peptide derived from Human Phospho-AKT1 (Ser473)
Immunogen Species
Homo sapiens (Human)
Conjugate
Non-conjugated
Clonality
Monoclonal
Isotype
Rabbit IgG
Clone No.
4H12
Purification Method
Affinity-chromatography
Concentration
It differs from different batches. Please contact us to confirm it.
Buffer
Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.
Form
Liquid
Tested Applications
ELISA, WB, IHC, IF, IP
Recommended Dilution
Application Recommended Dilution
WB 1:500-1:5000
IHC 1:50-1:200
IF 1:20-1:200
IP 1:200-1:1000
Protocols
ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) Protocol
Immunoprecipitation (IP) Protocol
Troubleshooting and FAQs
Antibody FAQs
Storage
Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
Lead Time
Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Description

AKT1, also known as protein kinase B, serves as a central node in the PI3K/AKT signaling pathway, regulating critical cellular processes including survival, proliferation, and metabolism. Phosphorylation at Serine 473 represents a key activation event that enables AKT1 to phosphorylate downstream substrates, making this modification an essential biomarker for studying pathway activation in cancer biology, metabolic disorders, and cell signaling research.

This recombinant monoclonal antibody, generated from clone 4H12, offers the reproducibility and consistency that phospho-specific detection demands. Because recombinant production ensures a sequence-defined binding region, you can expect reliable performance across experiments and between reagent lots—particularly valuable when tracking subtle changes in phosphorylation status over time or across treatment conditions.

Validation data demonstrates robust performance across multiple applications. Western blot analysis of 293 cell lysates treated with Calyculin A, a phosphatase inhibitor, confirms specific detection of phospho-AKT1 at the expected 60 kDa molecular weight, with clear signal enhancement in treated versus untreated samples. Immunohistochemistry staining has been validated in paraffin-embedded human breast cancer and lung cancer tissues, revealing the phosphorylation patterns relevant to oncology research. Immunofluorescence studies using EGF-stimulated HeLa cells demonstrate the antibody's utility for visualizing dynamic pathway activation at the cellular level. Additionally, immunoprecipitation capability has been confirmed in Calyculin A-treated 293 lysates, enabling enrichment studies and protein interaction analyses.

Whether investigating AKT pathway dysregulation in tumor samples, screening kinase inhibitor efficacy, or characterizing growth factor signaling dynamics, this antibody provides the phospho-specific detection and application flexibility essential for signal transduction research.

Usage
For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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Target Background

Function
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. Phosphorylates palladin (PALLD), modulating cytoskeletal organization and cell motility. Phosphorylates prohibitin (PHB), playing an important role in cell metabolism and proliferation. Phosphorylates 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. Phosphorylates PCK1 at 'Ser-90', reducing the binding affinity of PCK1 to oxaloacetate and changing PCK1 into an atypical protein kinase activity using GTP as donor. Also acts as an activator of TMEM175 potassium channel activity in response to growth factors: forms the lysoK(GF) complex together with TMEM175 and acts by promoting TMEM175 channel activation, independently of its protein kinase activity.
Gene References into Functions
  1. The optimal melatonin concentration (3 mM) significantly decreased the intracellular reactive oxygen species levels, caspase-3 activity and the percentage of both dead and apoptotic-like sperm cells and increased the vitality, progressive motility and total motility and AKT phosphorylation compared with the control group. PMID: 29196809
  2. The findings indicate that SPRY4 and SPRY4-IT1 may act as oncogenes in testicular germ cell tumors via activation of the PI3K/Akt signaling pathway. PMID: 29410498
  3. Results suggest that transient receptor potential vanilloid 4 (TRPV4) accelerates glioma migration and invasion through the AKT/Rac1 signaling, and TRPV4 might be considered as a potential target for glioma therapy. PMID: 29928875
  4. Data indicate a regulatory mechanism underlying drug resistance and suggest that tribbles homologue 2 (TRIB2) functions as a regulatory component of the PI3K network, activating AKT in cancer cells. PMID: 28276427
  5. findings indicated that shikonin inhibits proliferation and promotes apoptosis in human endometrioid endometrial cancer (EEC) cells by modulating the miR-106b/PTEN/AKT/mTOR signaling pathway, suggesting shikonin could act a potential therapeutic agent in the EEC treatment. PMID: 29449346
  6. SIRT6 inhibited proliferation, migration, and invasion of colon cancer cells by up-regulating PTEN expression and down-regulating AKT1 expression. PMID: 29957460
  7. LHPP suppresses cell proliferation and metastasis in cervical cancer, and promotes apoptosis by suppressing AKT activation. PMID: 29944886
  8. Data show that activated proto-oncogene protein Akt (AKT) directly phosphorylates Fas associated factor 1 (FAF1) reduces FAF1 at the plasma membrane and results in an increase in TGF-beta type II receptor (TbetaRII) at the cell surface. PMID: 28443643
  9. Data show while overexpression of AKT serine/threonine kinase 1 (AKT1) promoted local tumor growth, downregulation of AKT1 or overexpression of AKT serine/threonine kinase 2 (AKT2) promoted peritumoral invasion and lung metastasis. PMID: 28287129
  10. High AKT1 expression is associated with metastasis in ovarian cancer. PMID: 29739299
  11. Circ-CFH promotes glioma progression by sponging miR-149 and regulating the AKT1 signaling pathway. PMID: 30111766
  12. High AKT1 expression is associated with metastasis via epithelialmesenchymal transition carcinoma in colorectal cancer. PMID: 30066935
  13. High AKT1 expression is associated with tumor-node-metastasis in nonsmall cell lung cancer. PMID: 30106450
  14. High expression of AKT1 is associated with drug resistance and proliferation of breast cancer. PMID: 28165066
  15. Germline variants in AKT1 gene are associated with prostate cancer. PMID: 29298992
  16. High AKT1 expression is associated with cisplatinresistant oral cancer. PMID: 29956797
  17. that Akt1 was a novel target for miR-637, and its knockdown also induced cell growth inhibition and apoptosis in pancreatic ductal adenocarcinoma cells PMID: 29366808
  18. High AKT1 expression is associated with periodontitis. PMID: 30218719
  19. High AKT1 expression is associated with angiogenesis of esophageal squamous cell carcinoma. PMID: 30015941
  20. High AKT1 expression is associated with Pancreatic Ductal Adenocarcinoma Metastasis. PMID: 29386088
  21. In MCF-7 cells, AIB1 overexpression increases p-AKT (Ser 473) activity.In both T47D and MCF-7 cells overexpressing A1B1, p-AKT (Ser 473) expression was significantly increased in the presence or absence of IGF-1,but increased more in presence of IGF-1. PMID: 29808803
  22. In this study, we used the Ion Personal Genome Machine (PGM) and Ion Torrent Ampliseq Cancer panel to sequence hotspot regions from PIK3CA, AKT and PTEN genes to identify genetic mutations in 39 samples of TNBC subtype from Moroccan patients and to correlate the results with clinical-pathologic data PMID: 30227836
  23. AKT pathway is activated by CBX8 in in hepatocellular carcinoma. PMID: 29066512
  24. Here the authors identified a direct interaction of both MEK1 and MEK2 with AKT. The interaction between MEK and AKT affects cell migration and adhesion, but not proliferation. The specific mechanism of action of the MEK-AKT complex involves phosphorylation of the migration-related transcription factor FoxO1. PMID: 28225038
  25. miR-195 suppresses cell proliferation of ovarian cancer cells through regulation of VEGFR2 and AKT signaling pathways. PMID: 29845300
  26. High AKT1 expression is associated with cell growth, aggressiveness, metastasis in gastric cancer. PMID: 30015981
  27. this is the first report showing long-duration exposure to nicotine causes increased proliferation of human kidney epithelial cells through activation of AKT pathway. PMID: 29396723
  28. RBAP48 overexpression contributes to the radiosensitivity of AGS gastric cancer cells via phosphoinositide3kinase/protein kinase B pathway suppression. PMID: 29901205
  29. Activating Akt1 mutations alter DNA double strand break repair and radiosensitivity. PMID: 28209968
  30. PI3K-Akt pathway inhibitors, Akti-1/2 and LY294002, reduced PFKFB3 gene induction by PHA, as well as Fru-2,6-P2 and lactate production. Moreover, both inhibitors blocked activation and proliferation in response to PHA, showing the importance of PI3K/Akt signaling pathway in the antigen response of T-lymphocytes. PMID: 29435871
  31. RIO kinase 3 (RIOK3) positively regulates the activity of the AKT/mTOR pathway in glioma cells. PMID: 29233656
  32. High AKT1 phosphorylation is associated with colorectal carcinoma. PMID: 29970694
  33. Results show that AKT1 was associated with hypertension in Mexican Mestizos but not Mexican Amerindians. PMID: 30176313
  34. TERT could induce thyroid carcinoma cell proliferation mainly through the PTEN/AKT signaling pathway. PMID: 29901196
  35. findings uncover a new function of p53 in the regulation of Akt signaling and reveal how p53, ASS1, and Akt are interrelated to each other. PMID: 28560349
  36. We performed quantitative mass spectrometry of IAV1918-infected cells to measure host protein dysregulation. Selected proteins were validated by immunoblotting and phosphorylation levels of members of the PI3K/AKT/mTOR pathway were assessed. PMID: 29866590
  37. Radiation resistance tumors have upregulated Onzin and POU5F1 expression. PMID: 29596836
  38. The essential role of AKT in the endocrine therapy resistance in estrogen receptor-positive, HER2-negative breast cancer.[review] PMID: 29086897
  39. FAL1 may work as a ceRNA to modulate AKT1 expression via competitively binding to miR-637 in HSCR. PMID: 30062828
  40. The overexpression of CHIP significantly increased the migration and invasion of the DU145 cells, which is possible due to activation of the AKT signaling pathway and upregulation of vimentin. The expression level of CHIP was observed to be increased in human prostate cancer tissues compared with the adjacent normal tissue. PMID: 29693147
  41. Genistein (GE) inhibited the growth of human Cholangiocarcinoma (CCA) cell lines by reducing the activation of EGFR and AKT, and by attenuating the production of IL6. E2 and ER were also involved in the growth-inhibitory effect of GE in CCA cells. PMID: 29693152
  42. this study identifies ORP2 as a new regulatory nexus of Akt signaling, cellular energy metabolism, actin cytoskeletal function, cell migration, and proliferation. PMID: 29947926
  43. The role of USP18 in breast cancer provides a novel insight into the clinical application of the USP18/AKT/Skp2 pathway. PMID: 29749454
  44. Collectively, these results indicate that COX-1/PGE2/EP4 upregulates the beta-arr1 mediated Akt signaling pathway to provide mucosal protection in colitis. PMID: 28432343
  45. AKT kinase pathway is regulated by SPC24 in breast cancer. PMID: 30180968
  46. CREBRF promotes the proliferation of human gastric cancer cells via the AKT signaling pathway. PMID: 29729692
  47. these results indicate that miR124 transection inhibits the growth and aggressive of osteosarcoma, potentially via suppression of TGFbetamediated AKT/GSK3beta/snail family transcriptional repressor 1 (SNAIL1) signaling, suggesting miR124 may be a potential anticancer agent/target for osteosarcoma therapy. PMID: 29488603
  48. piperine reduced the expression of pAkt, MMP9 and pmTOR. Together, these data indicated that piperine may serve as a promising novel therapeutic agent to better overcome prostate cancer metastasis. PMID: 29488612
  49. S100A8 gene knockdown reduced cell proliferation in the HEC-1A cells compared with control cells, induced cell apoptosis, inhibited the phosphorylation of protein kinase B (Akt), and induced the expression of pro-apoptotic genes. PMID: 29595187
  50. intact keratin filaments are regulators for PKB/Akt and p44/42 activity, basal and in response to stretch. PMID: 29198699

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Involvement in disease
Breast cancer (BC); Colorectal cancer (CRC); Proteus syndrome (PROTEUSS); Cowden syndrome 6 (CWS6)
Subcellular Location
Cytoplasm. Nucleus. Cell membrane.
Protein Families
Protein kinase superfamily, AGC Ser/Thr protein kinase family, RAC subfamily
Tissue Specificity
Expressed in prostate cancer and levels increase from the normal to the malignant state (at protein level). Expressed in all human cell types so far analyzed. The Tyr-176 phosphorylated form shows a significant increase in expression in breast cancers dur
Database Links

HGNC: 391

OMIM: 114480

KEGG: hsa:207

STRING: 9606.ENSP00000270202

UniGene: Hs.525622

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Related pathways
cAMP signaling pathway Chemokine signaling pathway ErbB signaling pathway Estrogen signaling pathway HIF-1 signaling pathway Jak-STAT signaling pathway MAPK signaling pathway mTOR signaling pathway PI3K-Akt signaling pathway VEGF signaling pathway Focal adhesion Lipids and Inflammation in Atherogenesis TNF signaling pathway Apoptosis Autophagy Cellular senescence Ras signaling pathway Signaling pathways regulating pluripotency of stem cells B cell receptor signaling pathway Fc epsilon RI signaling pathway Fc gamma R-mediated phagocytosis Osteoclast differentiation T cell receptor signaling pathway Toll-like receptor signaling pathway Adipocytokine signaling pathway AGE-RAGE signaling pathway in diabetic complications AMPK signaling Apelin signaling pathway cGMP-PKG signaling pathway FoxO signaling pathway Glucagon signaling pathway Insulin signaling pathway Oxidative Phosphorylation Phospholipase D signaling pathway Rap1 signaling pathway Neurotrophin signaling pathway Target Therapy
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