Invitrogen™ Human AKT (Phospho) [pS473] ELISA Kit

ELISA

Supplier:  Invitrogen™ KHO0111

Catalog No. KHO0111

Description

The Human AKT (Phospho) [pS473] ELISA Kit is designed to detect and quantify the level of AKT (Phospho) [pS473] in fresh or frozen human cell lysates. The assay recognizes both natural and recombinant AKT (Phospho) [pS473]. Principle of the method A monoclonal capture antibody specific for AKT (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.

AKT1 (PKB alpha) is a serine/threonine kinase that regulates cell survival. The activated enzyme inhibits apoptosis and stimulates cell cycle progression by phosphorylating numerous targets in various cell types, including cancer cells. This protein kinase is activated by insulin, PI3K, IGF1 and various other growth and survival factors. Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including forkhead transcription factors, and caspase-9. The AKT pathway is a major target for cancer drug discovery.

Specifications

• Accession Number: P31749
• Assay Sensitivity: <0.8 U/mL
• Conjugate: HRP
• Sample Type: Cell Lysates
• Target Species: Rat, Mouse, Human
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene Symbol: AKT,CWS6,PKB,PKB-ALPHA,PRKBA,RAC,RAC-ALPHA,HIHGHH,PKBB,PKBBETA,PRKBB,RAC-BETA,MPPH,MPPH2,PKB-GAMMA,PKBG,PRKBG,RAC-Pk.-gamma,RAC-gamma,STK-2
• Interassay CV: 8.3%
• 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: Akt
• Regulatory Status: RUO
• Storage Requirements: 2°C to 8°C
• Test Time: 1 hrs 20 mins

• Assay Range: 1.6 to 100 U/mL
• Concentration: 1.6 U/mL
• Product Type: ELISA
• Substrate Type: Serine/Threonine Kinases
• For Use With (Application): Cell Analysis, Cell Signaling, Cell Viability, Proliferation and Function, Drug Discovery and Development, Immunoassays, Kinase Antibodies and Immunoassays, Kinase Biology, Ready-To-Use Immunoassay, Target and Lead Identification and Validation
• Gene ID (Entrez): 207
• Incubation Time: 4 hr.
• Intraassay CV: 6.9
• Label or Dye: HRP
• Quantity: 96 Tests
• Sample Volume: Cell Lysate 10 μL
• Target: AKT1
• Total Assay Time: 4 hrs

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov