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25% off all ELISA kits

Use code ELISA25-262 for 25% off all ELISAs through November.

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.

Product Description

Specifications

Size	1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Estimated Lead Time	1-2 business days
Species	Human, Mouse, Rat
Specificity	The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Akt (pSer473).
Compatible Sample Types	Tissue 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-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Design Principle	Sandwich-based
Method Of Detection	Colorimetric
Research Area	Angiogenesis, Apoptosis, Post-Translational Modifications, Inflammation, Phosphorylation, mTOR Signaling, PI3K-AKT Signaling, Insulin Signaling, Autophagy, Akt Signaling
Shipping Type	Blue 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

Prepare all reagents and samples as instructed in the manual.
Add 100 µl of sample or positive control to each well.
Incubate 2.5 h at RT or O/N at 4 °C.
Add 100 µl of prepared primary antibody to each well.
Incubate 1 h at RT.
Add 100 µl of prepared 1X HRP-Streptavidin to each well.
Incubate 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
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.

Have this product in hand?

Get your lot-specific datasheet at Ray's Docs. The datasheet linked below is our current version and may not match your lot.

Datasheet (PEL-AKT-S473-T.pdf)

Datasheet (PEL-AKT-S473.pdf)

SDS (PEL-AKT-S473-T.pdf)

SDS (PEL-AKT-S473.pdf)

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

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

Storage/Stability

Great product
Rating
This ELISA worked as well as expected. No complaints here!
Review by Lance, from Columbia University Medical Center, on Jan 13, 2020
Great product
Rating
This is a highly effective kit to semi-quantitatively measure phosphorylated Akt (Ser473) in lysates in a a 96-well plate format. Easy to read and quite reproducible. I would highly recommend it.
Review by Vinayak, from Moffitt Cancer Center, on Dec 8, 2019
Great product
Rating
Works well for detection in bone marrow stromal cell lysates
Review by William, on Apr 10, 2018

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Phospho-AKT (S473) ELISA Kit