Anti-Phospho-AKT1 (T450) Rabbit Monoclonal Antibody

SKU P00024-2
Size 100ug/vial
Reactivity Human, Mouse, Rat
Clonality Monoclonal
Clone Number HEA-1
Host Rabbit
Applications IP, WB

Overview

Product Name Anti-Phospho-AKT1 (T450) Rabbit Monoclonal Antibody
SKU/Catalog Number P00024-2
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.
Size 100ug/vial
Description Anti-Phospho-AKT1 (T450) Rabbit Monoclonal Antibodytested for IP, IHC, WB in Human, Mouse, Rat
Cite This Product Anti-Phospho-AKT1 (T450) Rabbit Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # P00024-2)
Host Rabbit
Contents/Buffer Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Form Liquid
Immunogen A synthesized peptide derived from human AKT1
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: 0.5-1mg/ml, actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
WB 1:1000-1:2000
IHC 1:50-1:200
IP 1:50

Images And Assay Conditions

Western blot analysis of Phospho-AKT1 (T450) expression in (1) 3T3 cell lysate treated with Lambda Phosphatase; (2) 3T3 cell lysate (P00024-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-AKT1 monoclonal antibody (Catalog # P00024-2) overnight at 4℃, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for AKT1

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P31749
Gene Name AKT1
Protein Name RAC-alpha serine/threonine-protein kinase
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 during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages. .
Alternative Names RAC-alpha serine/threonine-protein kinase;2.7.11.1;Protein kinase B;PKB;Protein kinase B alpha;PKB alpha;Proto-oncogene c-Akt;RAC-PK-alpha;AKT1;PKB, RAC;
Subcellular Localization Cytoplasm. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus.
Molecular Weight 55686 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein 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.
Research Areas Human, Mouse, Rat

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Monoclonal antibody for AKT1 detection. Host: Rabbit.Size: 100ug/vial. Tested applications: IP, IHC, WB. Reactive species: Human, Mouse, Rat AKT1 information: Molecular Weight: 55686 MW; Subcellular Localization: Cytoplasm. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus; 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 during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages.
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In stock
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P00024-2
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$299.00

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Publications

miR-21 Reduces Hydrogen Peroxide-Induced Apoptosis in c-kit
miR-21 increases c-kit cardiac stem cell proliferation in vitro through PTEN/PI3K/Akt signaling
A small interfering RNA targeting osteopontin as gastric cancer therapeutics
Short hairpin RNA targeting AKT1 and PI3K/p85 suppresses the proliferation and self-renewal of lung cancer stem cells
Protective Effects of Sodium (?)-5-Bromo-2-(%u03B1-Hydroxypentyl) Benzoate in a Rodent Model of Global Cerebral Ischemia
Vibration exercise decreases insulin resistance and modulates the insulin signaling pathway in a type 2 diabetic rat model
Epigallocatechin-3-Gallate Promotes the Growth of Mink Hair Follicles Through Sonic Hedgehog and Protein Kinase B Signaling Pathways
Hyperthermia induced HIF-1a expression of lung cancer through AKT and ERK signaling pathways
Epicatechin Plus Treadmill Exercise are Neuroprotective Against Moderate-stage Amyloid Precursor Protein/Presenilin 1 Mice
Progesterone alleviates hypoxic-ischemic brain injury via the Akt/GSK-3? signaling pathway
Li R, Cui K, Liu K, Li H, Zhang Y, Liu X, Chen R, Li M, Wang T, Wang S, Liu J, Rao K. Sci Rep. 2017 Oct 18;7(1):13464. doi: 10.1038/s41598-017-12907-1. Metabolic syndrome in rats is associated with erectile dysfunction by impairing PI3K/Akt/eNOS a...
Wang Q, Sun X, Li X, Dong X, Li P, Zhao L. Mol Med Rep. 2015 Jan;11(1):151-8. Doi: 10.3892/Mmr.2014.2762. Epub 2014 Oct 23. Resveratrol Attenuates Intermittent Hypoxia-Induced Insulin Resistance In Rats: Involvement Of Sirtuin 1 And The Phosphatid...
Wan J, Che Y, Kang N, Wu W. Mol Med Rep. 2015 Jul;12(1):83-92. Doi: 10.3892/Mmr.2015.3368. Epub 2015 Feb 17. Socs3 Blocks Hif-1?? Expression To Inhibit Proliferation And Angiogenesis Of Human Small Cell Lung Cancer By Downregulating Activation Of ...
Qin Q, Niu J, Wang Z, Xu W, Qiao Z, Gu Y. Cardiovasc Diabetol. 2013 Feb 26;12:37. Doi: 10.1186/1475-2840-12-37. Heparanase Induced By Advanced Glycation End Products (Ages) Promotes Macrophage Migration Involving Rage And Pi3K/Akt Pathway.
Wang Gg, Li W, Lu Xh, Zhao X, Xu L. Croat Med J. 2013 Apr;54(2):171-9. Taurine Attenuates Oxidative Stress And Alleviates Cardiac Failure In Type I Diabetic Rats.
Tang Y, Zhang X, Qi F, Chen M, Li Y, Liu L, He W, Li Z, Zu X. Exp Ther Med. 2015 May;9(5):1851-1856. Epub 2015 Feb 25. Afatinib Inhibits Proliferation And Invasion And Promotes Apoptosis Of The T24 Bladder Cancer Cell Line.
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