Custom Human and Mouse Phospho-4E-BP1 (Thr36) and Total 4E-BP1 Cell-Based ELISA

$633.00

RayBio® Human and Mouse Phospho-4E-BP1 (Thr36) and Total 4E-BP1 Cell-Based ELISA Kit. This assay semi-quantitatively measures 4E-BP1 phosphorylated at Threonine-36 as well as total 4E-BP1 in adherent cell lines.
Lead time 4-6 weeks

CBEL-4EBP1-T36-1
+

Antigen Information

Accession Number:
  • Q13541
  • Q60876
Gene ID:
  • 1978
Gene Symbol:
  • EIF4EBP1
Protein Name & Synonyms:
Eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) (eIF4E-binding protein 1) (Phosphorylated heat- and acid-stable protein regulated by insulin 1) (PHAS-I)
Target Species:
  • Human
  • Mouse

Assay Format

Pathway:
  • Akt Signaling
  • AMPK Signaling
  • HER/ErbB Signaling
  • IGF Signaling
  • mTOR Signaling
  • PI3K-AKT Signaling
Specificity:
This ELISA kit recognizes Human
Number of Targets Detected:
2
Species Detected:
  • Human
  • Mouse
Compatible Sample Types:
  • Adherent Cell Culture
Design Principle:
  • Cell-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Semi-Quantitative
Solid Support:
96-well Microplate

Product Specifications

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

Product Features

  • Rapidly measure phosphorylated protein in adherent cell lines
  • Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
  • No sample lysis is needed
  • Compatible with a standard ELISA plate reader

Application Notes

Kit Components
  • Uncoated 96-well Strip Microplate
  • Wash Buffers
  • 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
  1. Seed 10,000-30,000 cells into each well and incubate overnight.
  2. Apply various treatment, inhibitors or activators according to manufacturer's instructions.
  3. Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
  4. Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
  5. Add 200 µl of Blocking Solution and incubate for 1 h at 37°C.
  6. 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.
  7. Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 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.

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.

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