Human/Mouse/Rat Phospho-AKT (S473) and Total Akt ELISA

$536.00
PEL-AKT-S473-T-1

RayBio® Human/Mouse/Rat Phospho-Akt (S473) and Total Akt ELISA Kit. This assay semi-quantitatively measures phosphorylated Akt (Ser473) and Total Akt in lysate samples.

Product Features

  • Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
  • Screen numerous different cell lysates without performing a Western Blot analysis
  • Minimal hands-on time, convenient, and non-radioactive material

Specificity

The antibody pair provided in this kit recognizes human, mouse and rat Akt phosphorylated at Serine-473 + pan Akt.

Target Protein Information

Synonyms
RAC-alpha serine/threonine-protein kinase (EC 2.7.11.1) (Protein kinase B) (PKB) (Protein kinase B alpha) (PKB alpha) (Proto-oncogene c-Akt) (RAC-PK-alpha)
Accession Number
Gene Names
AKT1 PKB RAC
Gene ID
207

Species Detected

Human, Mouse, Rat

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Anti-Phospho Antibody
  • Anti-Pan Antibody
  • HRP-Conjugated Secondary Antibody
  • Streptavidin-Conjugated HRP
  • Assay Diluent
  • TMB One-Step Substrate
  • Stop Solution
  • Lysis Buffer
  • Positive Control Sample
Other Materials Required
  • Distilled or deionized water
  • 100 ml and 1 liter graduated cylinders
  • Tubes to prepare sample dilutions
  • Protease and Phosphatase inhibitors
  • Precision pipettes to deliver 2 µl to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
  • Benchtop rocker or shaker
  • Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
  1. Prepare all reagents and samples as instructed in the manual.
  2. Add 100 µl of sample or positive control to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 µl of prepared primary antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.
  7. Incubate 1 h at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Storage/Stability

Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

Antigen Information

Gene Symbol:
  • AKT1

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Array/ELISA Format

Pathway:
  • Insulin Signaling
  • mTOR Signaling
  • PI3K-AKT Signaling
Species Detected:
  • Human
  • Mouse
  • Rat
Solid Support:
96-well Microplate
Design Principle:
  • Sandwich-based
Quantitative/Semi-Quantitative:
Semi-Quantitative
Method of Detection:
Colorimetric
Compatible Sample Types:
  • Cell Lysates
  • Tissue Lysates
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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.