ELISA/assay

Rat Protein Kinase B, PKB/Akt ELISA Kit

MBS701882

48-Strip-Wells

590 USD

ELISA Kit

Rat Protein Kinase B, PKB/Akt ELISA Kit

v-akt murine thymoma viral oncogene homolog 1

Rat Protein Kinase B (PKB/Akt) ELISA Kit; AKT; MGC99656; PKB; PKB-ALPHA; PRKBA; RAC; RAC-ALPHA; AKT1 kinase; RAC-alpha serine/threonine-protein kinase; protein kinase B; rac protein kinase alpha; v-akt murine thymoma viral oncogene homolog 1

RAC-alpha serine/threonine-protein kinase; RAC-alpha serine/threonine-protein kinase; RAC-alpha serine/threonine-protein kinase; PKB alpha; AKT1 kinase; RAC-PK-alpha; protein kinase B alpha; thymoma viral proto-oncogene 1; RAC protein kinase alpha RAC-PK alpha; murine thymoma viral (v-akt) oncogene homolog 1; v-akt murine thymoma viral oncogene homolog 1; Protein kinase B; PKB; Protein kinase B alpha; PKB alpha; RAC-PK-alpha

AKT1

Akt1; Akt1; Akt; PKB; PKB alpha

AKT1_RAT

Rat

480

This assay has high sensitivity and excellent specificity for detection of Rat AKT1. No significant cross-reactivity or interference between Rat AKT1 and analogues was observed.

Unopened test kits should be stored at 2 to 8 degree C upon receipt. Please refer to pdf manual for further storage instructions.

Samples: Serum, plasma, cell culture supernates, tissue homogenates. Detection Range: 0.156 ng/ml-10 ng/ml. Sensitivity: 0.039 ng/ml

Intra-assay Precision: Intra-assay Precision (Precision within an assay): CV% is less than 8%. Three samples of known concentration were tested twenty times on one plate to assess. Inter-assay Precision (Precision between assays): CV% is less than 10%. Three samples of known concentration were tested in twenty assays to assess. Detection Wavelength: 450 nm. Sample Volume: 50-100ul. Protein Biological Process 1: Apoptosis/Autophagy. Protein Biological Process 3: Apoptosis

Introduction: Protein kinase B or Akt (PKB/Akt) is a serine/threonine kinase, which in mammals comprises three highly homologous members known as PKBalpha (Akt1), PKBbeta (Akt2), and PKBgamma (Akt3). PKB/Akt is activated in cells exposed to diverse stimuli such as hormones, growth factors, and extracellular matrix components. The activation mechanism remains to be fully characterised but occurs downstream of phosphoinositide 3-kinase (PI-3K). PI-3K generates phosphatidylinositol-3,4,5-trisphosphate (PIP(3)), a lipid second messenger essential for the translocation of PKB/Akt to the plasma membrane where it is phosphorylated and activated by phosphoinositide-dependent kinase-1 (PDK-1) and possibly other kinases. PKB/Akt phosphorylates and regulates the function of many cellular proteins involved in processes that include metabolism, apoptosis, and proliferation. Recent evidence indicates that PKB/Akt is frequently constitutively active in many types of human cancer. Constitutive PKB/Akt activation can occur due to amplification of PKB/Akt genes or as a result of mutations in components of the signalling pathway that activates PKB/Akt. Although the mechanisms have not yet been fully characterised, constitutive PKB/Akt signalling is believed to promote proliferation and increased cell survival and thereby contributing to cancer progression. This review surveys recent developments in understanding the mechanisms and consequences of PKB/Akt activation in human malignancy. Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to PKB. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for PKB. and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain PKB, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of PKB. in the samples is then determined by comparing the O.D. of the samples to the standard curve.

15100164

NP_150233.1

NM_033230.2

P47196

55,735 Da

AKT Phosphorylates Targets In The Cytosol Pathway (1010029); AKT Phosphorylates Targets In The Nucleus Pathway (1010030); AKT-mediated Inactivation Of FOXO1A Pathway (1009917); AMPK Signaling Pathway (989832); AMPK Signaling Pathway (992181); Activation Of BAD And Translocation To Mitochondria Pathway (1010642); Activation Of BH3-only Proteins Pathway (1010641); Acute Myeloid Leukemia Pathway (83505); Acute Myeloid Leukemia Pathway (529); Adaptive Immune System Pathway (1010925)

inhibits JUN kinase activation and mediates inhibition of apoptosis [RGD, Feb 2006]

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 PI3P-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 (PI3K) 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 4 PublicationsManual assertion based on experiment in:Ref.2. Catalytic activity: ATP + a protein = ADP + a phosphoprotein. Enzyme regulation: Three specific sites, one in the kinase domain (Thr-308) and the two other ones in the C-terminal regulatory region (Ser-473 and Tyr-474), need to be phosphorylated for its full activation.

48-Strip-Wells

590 USD

96-Strip-Wells

850

5x96-Strip-Wells

2880

10x96-Strip-Wells

5440