LANCE Ultra phospho-AKT1/2/3 (Ser473) kits are designed for the detection of AKT1, AKT2, and AKT3 (phosphorylated at residue S473) in cell lysates using a simple, homogeneous LANCE Ultra assay (no wash steps). This assay is compatible with both adherent and suspension cells.
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For research use only. Not for use in diagnostic procedures.
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Please note control lysates are NOT included in kits. Control lysates are sold separately, catalog number TRF4002S.
LANCE® and LANCE® (Lanthanide chelate excite) Ultra are homogeneous (no wash) TR-FRET (time-resolved fluorescence resonance energy transfer) technologies. One antibody of interest is labeled with a donor fluorophore (a LANCE Europium chelate) and the second antibody is labeled with an acceptor fluorophore [ULight™ dye]. Upon excitation at 320 or 340 nm, energy can be transferred from the donor Europium chelate to the acceptor fluorophore if sufficiently close for FRET (~10 nm). This results in the emission of light at 665 nm. Data are represented as ratiometric (665/615 nm X 10,000).
AKT, also known as protein kinase B, is an important regulator of numerous cellular processes including cell survival, glucose metabolism, transcription, and apoptosis. AKT has three closely related isoforms (AKT1, AKT2, and AKT3), and these isoforms have been shown to act on both common and unique downstream substrates. AKT activation can be induced by a number of stimuli, whereby AKT is transported to membrane for phosphorylation. Activation occurs upon AKT phosphorylation of Thr308 by PDK1 and of Ser473 by mTORC2. AKT subsequently dissociates from the membrane and phosphorylates targets both in the cytoplasm, as well as, the cell nucleus. Deregulation of AKT has been implicated in a number of disease states including cancer, cardiovascular disease, and diabetes.
|Assay Target Class||Phosphoprotein|
|Detection Method||Time-Resolved Fluorescence (TRF), TR-FRET|
|Product Brand Name||LANCE Ultra|
|Shipping Condition||Blue Ice|
|Unit Size||500 Assay Points|
This application note describes a comparison of two TR-FRET based technologies for measuring phospho-ERK