Invitrogen™ AKT1 (Phospho) [pS473] Human ELISA Kit, Ultrasensitive

High Sensitivity Sandwich ELISA Kit

Brand:  Invitrogen™ KHO0541

Product Code. 10642644

Includes: AKT Antibody Coated 96-Well Plate, AKT [pS473] Standard, Standard Diluent Buffer, AKT [pS473] Detection Antibody, Anti-Rabbit IgG-HRP (100X), HRP Diluent, Wash Buffer Concentrate (25X), Stabilized Chromogen, TMB, Stop Solution, Plate Covers, Detailed protocol with validation tests

Description

Kit is designed to detect and quantify the level of AKT1 (Phospho) [pS473] in fresh or frozen human cell lysates. Cross-reactivity with mouse and rat cells has been observed. The assay recognizes both natural and recombinant AKT1 (Phospho) [pS473]. Principle of the method A monoclonal capture antibody specific for AKT1 (Phospho) [pS473] has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of target protein present in the original specimen and the optical density can be read on a standard microplate reader. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Mutations in this gene have been associated with the Proteus syndrome. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Jul 2011]

Order Info

Shipping condition: Wet ice

Specifications

• Accession Number: P31749,P31750,P47196
• Assay Sensitivity: 0.2 U/mL
• Conjugate: HRP
• Sample Type: Cell Lysates
• Target Species: Human, Mouse, Rat
• Gene ID (Entrez): 11651, 207, 24185
• Incubation Time: 4 hr.
• Intraassay CV: 4.7%
• Label or Dye: HRP
• Quantity: 96 Tests
• Sample Volume: Cell Lysate,10 μL
• Target: Human, Mouse, Rat
• Total Assay Time: 4 hr.

• Assay Range: 0.39-25 U/mL
• Concentration: 0.3 to 25U/mL
• Product Type: ELISA Kit
• Substrate Type: Serine/Threonine Kinases
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene Symbol: AKT,CWS6,PKB,PKB-ALPHA,PRKBA,RAC,RAC-ALPHA
• Interassay CV: 5.2%
• Kit Contents: Pre-coated 96 well plate, Standard, Standard Dilution Buffer, Detection Antibody, anti-rabbit-HRP, HRP Diluent, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Marker Gene: Akt1
• Regulatory Status: RUO
• Storage Requirements: 2°C to 8°C
• Test Time: 1 hr. 20 min.

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