Technical guide & protocol Tech Support

Product Specifications

Analytical sensitivity

0.2 U/mL

Assay range

0.39-25 U/mL

Sample type/volume

Cell Lysate
10 µL

Hands-on time

1 hr 20 min

Time-to-result

4 hr

Homogenous (no wash)

No

Interassay CV

5.2%

Intraassay CV

4.7%

Instrument

Colorimetric Microplate Reader

Product size

96 Tests

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

Shipping conditions

Wet ice

Storage

2-8°C

Protein name

AKT1

Target site-specificity

Phosphorylated Ser473

Protein subtype

Serine/Threonine Kinases

Protein family

Kinases & Inhibitors

Species (tested)

Human, Mouse, Rat

Assay kit format

High Sensitivity Sandwich ELISA Kit

Detector antibody conjugate

HRP

Label or dye

HRP

About This Kit

The Human AKT1 (Phospho) [pS473] ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) 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.

Target information

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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Gene aliases : AKT, AKT1, CWS6, PKB, PKB-ALPHA, PKB/Akt, PKBalpha, PRKBA, RAC, RAC-ALPHA

Gene ID : (Human) 207, (Mouse) 11651, (Rat) 24185

Gene symbol : AKT1, Akt1, Akt1

Protein Aliases : AKT1 kinase, AKT1m, Akt1m protein, murine thymoma viral (v-akt) oncogene homolog 1, PKB, PKB alpha, Protein kinase B, Protein kinase B alpha, protein kinase B-alpha, Proto-oncogene c-Akt, rac protein kinase alpha, RAC protein kinase alpha RAC-PK alpha, RAC-alpha serine/threonine-protein kinase, RAC-PK-alpha, related to A and C kinases, serine-threonine protein kinase, Thymoma viral proto-oncogene, thymoma viral proto-oncogene 1, v-akt murine thymoma viral oncogene homolog 1, v-akt murine thymoma viral oncogene-like protein 1

UniProt ID (Human) P31749, (Mouse) P31750, (Rat) P47196

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