Phospho-AKT (S473) ELISA Kit

Cat# PEL-AKT-S473-1

Size : 1PlateKit

Brand : RayBiotech

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Phospho-AKT (S473) ELISA Kit

The RayBiotech Phospho-AKT (S473) ELISA Kit measures phosphorylated AKT (Ser473) and total (pan) AKT in cell and tissue lysates.
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Product Description

Specifications

Size1 Plate Kit, 2 Plate Kit, 5 Plate Kit
SpeciesHuman, Mouse, Rat
Specificity
The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Akt (pSer473).
Compatible Sample TypesTissue Lysates, Cell Lysates
Accession Number
P31749
Gene Id
391
Gene Symbols
AKT1, PKB, RAC
Protein Name / 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)
Quantitative/Semi-QuantitativeSemi-Quantitative
Solid Support
96-well Microplate
Design PrincipleSandwich-based
Method Of DetectionColorimetric
Research AreaAngiogenesis, Apoptosis, Post-Translational Modifications, Inflammation, Phosphorylation, mTOR Signaling, PI3K-AKT Signaling, Insulin Signaling, Autophagy, Akt Signaling
Shipping TypeBlue ice
Storage-20°C

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

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Anti-Phospho Antibody
  • Anti-Pan Antibody (*not included in phospho-only version)
  • HRP-Conjugated Secondary Antibody
  • 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.

citations

  • Effect of curcumin-loaded polydopamine nanoparticles on the expression of AKT, HDAC6, and HSF1 in Trembler-J Schwann cell cultures

    Species:
    Mouse
    Sample Type:
    Cell Lysate (wt, TrJ schwann cell lysates); Tissue Lysate / Homogenate
    Year:
    2026
    Journal Name:
    Research Square
  • Novel insights into gut health: Cilostazol strengthens gut integrity by adjusting TLR-2/NF-κB/IL-23 and CD44/AKT/GSK-3β/cyclin-D1 trajectories in methotrexate-induced mucositis model

    Year:
    2024
    Journal Name:
    European Journal of Pharmacology
    PubMed ID:
    38795758
  • Local application of engineered insulin-like growth factor I mRNA demonstrates regenerative therapeutic potential in vivo

    Species:
    Mouse, Rat
    Sample Type:
    muscle tissue
    Year:
    2023
    Journal Name:
    Molecular Therapy-Nucleic Acids
  • Renoprotective Effect of Pitavastatin against TAA-Induced Renal Injury: Involvement of the miR-93/PTEN/AKT/mTOR Pathway

    Species:
    Rat
    Sample Type:
    Tissue Lysate / Homogenate (renal tissue homogenate)
    Year:
    2024
    Journal Name:
    Advances in pharmacological and pharmaceutical sciences
    PubMed ID:
    38293706
  • Dapagliflozin/Hesperidin Combination Mitigates Lipopolysaccharide-Induced Alzheimer’s Disease in Rats

    Species:
    Rat
    Sample Type:
    hippocampal
    Year:
    2023
    Journal Name:
    Pharmaceuticals
  • The Role of Beta-Hydroxybutyrate in Mitigating the Inflammatory and Metabolic Consequences of Uric Acid

    Species:
    Mouse
    Sample Type:
    Cell Lysate (cell lysates C2C12 murine muscle cells); Tissue Lysate / Homogenate
    Year:
    2024
    Journal Name:
    Metabolites
    PubMed ID:
    39728460
  • Empagliflozin enhances neuroplasticity in rotenone-induced parkinsonism: Role of BDNF, CREB and Npas4

    Species:
    Mouse
    Sample Type:
    brain
    Year:
    2022
    Journal Name:
    LIFE SCIENCES
  • Glimepiride mitigates tauopathy and neuroinflammation in P301S transgenic mice: role of AKT/GSK3β signaling

    Species:
    Mouse
    Sample Type:
    brain
    Year:
    2022
    Journal Name:
    INFLAMMOPHARMACOLOGY
  • Mitoquinone Alleviates Donation after Cardiac Death Kidney Injury during Hypothermic Machine Perfusion in Rat Model

    Species:
    Rat
    Sample Type:
    kidney tissue
    Year:
    2023
    Journal Name:
    International Journal of Molecular Sciences
  • Icosapent ethyl alleviates acetic acid-induced ulcerative colitis via modulation of SIRT1 signaling pathway in rats

    Species:
    Rat
    Sample Type:
    colon
    Year:
    2022
    Journal Name:
    International Immunopharmacology

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