Phospho-AKT1 (Ser473) Recombinant Monoclonal Antibody

說明書

中文名稱：

磷酸化-AKT1 (Ser473)重組抗體
貨號：

CSB-RA001553A473phHU
規格：

￥1320
圖片：
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)
其他：

產品詳情
產品評價
相關產品
靶點詳情

產品詳情

產品描述：

Phospho-AKT1 (Ser473) Recombinant Monoclonal Antibody（CSB-RA001553A473phHU）是一種高特異性抗體，靶向AKT1蛋白Ser473位點的磷酸化修飾。AKT1是PI3K/AKT/mTOR信號通路的核心激酶，其Ser473位點的磷酸化是激活下游細胞存活、增殖和代謝調控功能的關鍵步驟，在腫瘤發生、耐藥性及代謝性疾病研究中具有重要價值。該抗體通過重組技術制備，具有嚴格的批次一致性，經多種應用驗證：在蛋白質印跡（WB）中可清晰檢測AKT1磷酸化信號，推薦稀釋范圍1:500-1:5000；免疫組織化學（IHC）和免疫熒光（IF）分別支持1:50-1:200和1:20-1:200的稀釋比例，適用于石蠟切片或細胞樣本的定位分析；同時可在免疫沉淀（IP）中以1:200-1:1000的稀釋度高效富集磷酸化AKT1蛋白。其卓越的特異性已在人源、小鼠等物種樣本中得到驗證，適用于研究AKT信號通路的動態調控機制、腫瘤微環境中的異常激活現象以及藥物靶向干預效果評估，為癌癥生物學、代謝疾病及神經退行性病變等領域的分子機制研究提供可靠工具。
Uniprot No.：

P31749
基因名：

AKT1
別名：

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
反應種屬：

Human
免疫原：

A synthesized peptide derived from Human Phospho-AKT1 (Ser473)
免疫原種屬：

Homo sapiens (Human)
標記方式：

Non-conjugated
克隆類型：

Monoclonal
抗體亞型：

Rabbit IgG
純化方式：

Affinity-chromatography
克隆號：

4H12
濃度：

It differs from different batches. Please contact us to confirm it.
保存緩沖液：

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.
產品提供形式：

Liquid
應用范圍：

ELISA, WB, IHC, IF, IP

推薦稀釋比：

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
儲存條件：

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
貨期：

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.
用途：

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

產品評價

靶點詳情

功能：

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.
基因功能參考文獻：
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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相關疾病：

Breast cancer (BC); Colorectal cancer (CRC); Proteus syndrome (PROTEUSS); Cowden syndrome 6 (CWS6)
亞細胞定位：

Cytoplasm. Nucleus. Cell membrane.
蛋白家族：

Protein kinase superfamily, AGC Ser/Thr protein kinase family, RAC subfamily
組織特異性：

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
數據庫鏈接：

HGNC: 391

OMIM: 114480

KEGG: hsa:207

STRING: 9606.ENSP00000270202

UniGene: Hs.525622

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