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Human/Mouse/Rat Phospho-AKT (S473) PharmaGenie ELISA Kit (SBRS1743)

SKU:
SBRS1743
Product type:
ELISA Kit
Reactivity:
Human
Reactivity:
Mouse
Reactivity:
Rat
Application:
ELISA
ELISA Type:
Sandwich
€760
Frequently bought together:

Description

Human/Mouse/Rat Phospho-AKT (S473) PharmaGenie ELISA - Kit Data

Product SKU:SBRS1743
Size:96T
Application:The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Akt (pSer473).
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.

Kit Components:

1.Pre-Coated 96-well Strip Microplate
2.Wash Buffer
3.Anti-Phospho Antibody
4.HRP-Conjugated Secondary Antibody
5.Assay Diluent
6.TMB One-Step Substrate
7.Stop Solution
8.Lysis Buffer
9.Positive Control Sample

Other materials and equipment required:

The Assay Genie Human/Mouse/Rat Phospho-AKT (S473) ELISA Kit 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

Kit Protocol:

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.