Human/Mouse/Rat Phospho-AKT (S473) and Total Akt PharmaGenie ELISA Kit (SBRS1745)

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SKU:
SBRS1745
Product Type:
ELISA Kit
Reactivity:
Human
Reactivity:
Mouse
Reactivity:
Rat
Applications:
ELISA
ELISA Type:
Sandwich

Description

system_update_altDatasheet

Human/Mouse/Rat Phospho-AKT (S473) and Total Akt PharmaGenie ELISA Kit (SBRS1745)

Product SKU:SBRS1745
Size:96T
Application:The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Akt (pSer473) + pan Akt.
Uniprot:P31749
Gene ID:391
Gene Names:AKT1 / PKB / RAC
Pathway:Insulin Signaling / mTOR Signaling / PI3K-AKT Signaling
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)
Target Species:Human, Mouse, Rat
Compatible Sample Types:Cell Lysates, Tissue Lysates
Design Principle:Sandwich-based
Method of Detection:Colorimetric
Quantitative/Semi-Quantitative:Semi-Quantitative
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.
  1. Pre-Coated 96-well Strip Microplate
  2. Wash Buffer
  3. Anti-Phospho Antibody
  4. Anti-Pan Antibody
  5. HRP-Conjugated Secondary Antibody
  6. Streptavidin-Conjugated HRP
  7. Assay Diluent
  8. TMB One-Step Substrate
  9. Stop Solution
  10. Lysis Buffer
  11. Positive Control Sample

Other materials and equipment required:

The Assay Genie Human/Mouse/Rat Phospho-AKT (S473) and Total Akt PharmaGenie ELISA Kit (SBRS1745) will require other equipment and materials to carry out the assay. Please see list below for further details.

  • 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 ul 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
  1. Prepare all reagents and samples as instructed in the manual.
  2. Add 100 ul of sample or positive control to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 ul of prepared primary antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 ul of prepared 1X HRP-Streptavidin to each well.
  7. Incubate 1 h at RT.
  8. Add 100 ul of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 ul of Stop Solution to each well.
  11. Read at 450 nm immediately.
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