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Human/Mouse/Rat AKT (S473) Cell-Based ELISA Kit
Cell-Based Human/Mouse/Rat AKT (S473) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.
Product Description
Specifications
Size
1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species
Human, Mouse, Rat
Accession Number
P31749
Gene Id
207
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
Specificity
The antibodies provided in this kit recognizes human/mouse/rat AKT phosphorylated at Serine-473 and total AKT for comparison.
Compatible Sample Types
Adherent Cell Culture
Solid Support
96-well Microplate
Method Of Detection
Colorimetric
Design Principle
Cell-based
Research Area
Post-Translational Modifications, Phosphorylation, mTOR Signaling, PI3K-AKT Signaling, Insulin Signaling
Estimated Lead Time
1-2 business days
Shipping Type
Blue ice
Storage
-20°C
Risk-Free Guarantee
We offer a 100% guarantee on all ELISA kits and membrane cytokine arrays.
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$5 Amazon gift card in every kit box purchased.
Product Features Site and signal pathway-specific In vitro detection of adherent cell culture No sample lysis is needed Compatible with a standard ELISA plate reader Faster results than with ELISA Adaptable for high-throughput screening and drug discovery Application Notes Kit Components
uncoated 96-well Microplate Wash Buffer A Wash Buffer B Fixing Solution Quenching Buffer Blocking Buffer Anti-phospho antibody Anti-pan antibody HRP-Conjugated Secondary Antibody TMB One-Step Substrate Stop Solution 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
Seed 10,000-30,000 cells into each well and incubate overnight. Apply various treatment, inhibitors or activators according to manufacture's instructions. Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking. Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT. Add 200 µl of Blocking Solution and incubate for 1 h at 37 ° C. Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT. Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 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 The entire kit may be stored at -20° C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.
Great product
The kit was convenient and fast. The instructions are clear and the results are reproducible. Overall, it was a viable faster alternative than recreating all the component from individual reagents. Also, great costumer service.
Review by
Moises Guardado,
from
SUNY Stony Brook University,
on
Jun 11, 2018
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