Invitrogen™ Human AKT (Total) ELISA Kit

Description

ELISA

Kit is designed to detect and quantify the level of AKT (Total) in fresh or frozen human cell lysates. Cross-reactivity in mouse and rat cell lysates has been observed. The assay recognizes both natural and recomb. AKT (Total), independent of its phosphorylation state. Principle of the method A monoclonal capture antibody specific for AKT (Total) has been coated onto the wells of 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 HRP 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.

AKT also known as protein kinase B (PKB) or RAS-alpha, is an ubiquitous serine/threonine kinase that plays an important role in diverse biological responses such as regulation of metabolism, cell survival and growth by phosphorylating multiple proteins. 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,P31750,P31751,P47196,P47197,Q60823,Q63484,Q9WUA6,Q9Y243
• Assay Range: 0.3 to 20 ng/mL
• Assay Sensitivity: <0.1 ng/mL
• Concentration: 0.3 to 20 ng/mL
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Cell Lysates
• Substrate Type: Serine/Threonine Kinases
• Target Species: Mouse, Rat
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 10000, 11651, 11652, 207, 208, 23797, 24185, 25233, 29414
• 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
• Incubation Time: 4 hr.
• Interassay CV: 9.3%
• Intraassay CV: 7.7
• 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
• Label or Dye: HRP
• Marker Gene: Akt
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Cell Lysate 10 μL
• Storage Requirements: 2°C to 8°C
• Target: AKT Pan
• Test Time: 1 hrs 20 mins
• Total Assay Time: 4 hrs

Safety and Handling

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