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Human and Mouse Phospho-FOXO3 (S253) and Total FOXO3 ELISA

RayBio® Human and Mouse Phospho-FOXO3 (Ser253) and Total FOXO3 ELISA Kit. This assay semi-quantitatively measures FOXO3 phosphorylated at Serine-263 as well as total FOXO3 in lysate samples.

$2,164.00  
PEL-FOXO3-S253-T-5
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Antigen Information

Accession Number:
  • O43524
  • Q9WVH4
Gene ID:
  • 2309
Gene Symbols:
  • FKHRL1
  • FOXO3
  • FOXO3A
Protein Name & Synonyms:
Forkhead box protein O3 (AF6q21 protein) (Forkhead in rhabdomyosarcoma-like 1)
Target Species:
  • Human
  • Mouse

Assay Format

Specificity:
This ELISA kit recognizes Human

Product Specifications

Size:
1, 2, or 5 x 96-Well Microplate Kit

Product Features

  • Rapidly measure phosphorylated protein in lysates
  • 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
  • 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.

Typical Data

Positive Control
HeLa cells were treated with Calyculin A. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
Calyculin A Stimulation of HeLa Cell Lines
HeLa cells were treated or untreated with Calyculin A. Cell lysates were analyzed using this phosphoELISA and Western Blot.


Storage/Stability

Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.

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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.