Custom Human Phospho-APP (Thr668) and Total APP Cell-Based ELISA

RayBio® Human Phospho-APP (Thr668) and Total APP Cell-Based ELISA Kit. This assay semi-quantitatively measures APP phosphorylated at Threonine-668 as well as total APP in adherent cell lines.
Lead time 4-6 weeks

$633.00  
CBEL-APP-T668-1
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Antigen Information

Accession Number:
  • P05067
Gene ID:
  • 351
Gene Symbols:
  • A4
  • AD1
  • APP
Protein Name & Synonyms:
Amyloid beta A4 protein (ABPP) (APPI) (APP) (Alzheimer disease amyloid protein) (Cerebral vascular amyloid peptide) (CVAP) (PreA4) (Protease nexin-II) (PN-II) [Cleaved into: N-APP Soluble APP-alpha (S-APP-alpha) Soluble APP-beta (S-APP-beta) C99 Beta-amyloid protein 42 (Beta-APP42) Beta-amyloid protein 40 (Beta-APP40) C83 P3(42) P3(40) C80 Gamma-secretase C-terminal fragment 59 (Amyloid intracellular domain 59) (AICD-59) (AID(59)) (Gamma-CTF(59)) Gamma-secretase C-terminal fragment 57 (Amyloid intracellular domain 57) (AICD-57) (AID(57)) (Gamma-CTF(57)) Gamma-secretase C-terminal fragment 50 (Amyloid intracellular domain 50) (AICD-50) (AID(50)) (Gamma-CTF(50)) C31]
Target Species:
  • Human

Assay Format

Specificity:
This ELISA kit recognizes Human APP phosphorylated at site Threonine-668 as well as total APP.
Number of Targets Detected:
2
Species Detected:
  • Human
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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