AKT [pS473] Phospho-ELISA Kit, Human

Catalog number: KHO0111

Novex™  Related applications: Protein Assays and Analysis

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Description

The AKT [pS473] ELISA research use only kit is designed to detect and quantify the level of AKT protein phosphorylated at serine residue 473. This assay is intended for the detection of AKT [pS473] from human, mouse, or rat cell lysates using 96-well plates and a microplate reader.

Easy to run
More sensitive than westerns
Convenient precoated, removable 8-well strips

Easy-to-Run Sandwich ELISA
The total assay incubation time is only 4 hours. A monoclonal capture antibody specific for AKT has been coated onto the wells of the 96-well plate provided. During the first incubation, standards of known AKT [pS473] content and unknown samples are pipetted into the wells and the AKT antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for AKT phosphorylated at serine 473 is added to the wells. During the second incubation, this antibody serves as a detection antibody by binding to the immobilized AKT 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 AKT [pS473] present in the original specimen and the optical density can be read on a standard microplate reader.

Detect Low-Expressing Proteins and Use Less Sample
ELISA kits designed to measure intracellular signaling targets are 2–10 times more sensitive than western blotting. The improved sensitivity enables you to detect low-expressing proteins that otherwise may not be distinguishable from background. In addition, the amount of sample needed to run the assay is less than what is needed for western blots.

Ready-to-Use AKT [pS473] ELISA Kit Contains These Reagents, Buffers, and Supplies:
• AKT Antibody Coated 96-Well Plate
• AKT (Total) Standard
• Standard Diluent Buffer
• AKT (Total) 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

Convenient Kit Format
The kit comes with a capture antibody precoated to the plate, eliminating an overnight coating process. The plates are machine coated to provide low variability well-to-well. Furthermore, the plate comes as removable 8-well strips allowing you to run as few samples as you want. In addition, the standards provided in the kit can be used to get quantitative results off the standard curve and/or can be used as a positive control.

Rigorous Validation
Each ELISA kit is validated for sensitivity, specificity, precision, and lot-to-lot consistency. See protocol insert for more information on validation.

Part of the ELISA Kit Family
We have a wide menu of ELISA research use only kits. With excellent sensitivity, proven specificity, and consistent performance, these optimized ELISA kits offer you a choice when it comes to getting physiologically relevant and quantitative results.

For research use only. Not intended for human or animal therapeutic or diagnostic use.

Related Links:

Learn more about ELISA Kits
Learn more about other Immunoassays
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Protein Subtype: Serine⁄Threonine Kinases
Shipping Condition: Wet Ice
Marker: Akt
Gene ID: 10000, 207, 208
Species: Mouse, Human, Rat
Gene Symbol: AKT1, AKT3, AKT2
Sensitivity: <0.8 U⁄mL (4000 cells)
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 96 assays
Concentration: 1.6 U⁄ml
Sample Volume: 10 µl
Protein Family: Kinases & Inhibitors
Incubation Time: 4 hrs
Detection Method: Colorimetric
Research Category: Signal Transduction
Sample Type (Specific): Cell Extracts
For Use With (Equipment): Microplate Reader

Contents & storage

Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.

Documents

Manuals & protocols

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