Cell-Based Phosphorylation ELISA Kits

ELISA Genie Cell-Based Phosphorylation ELISA Kits

The ELISA Genie Cell-Based phospho-specific ELISA Kits can be used for measuring the relative amounts of phosphorylated protein in whole cultured cells, as well as screening for the effects that various treatments, inhibitors (ie. siRNA or chemicals), or activators have on protein phosphorylation.

These in vitro immunohistochemistry ELISA assays are a premium range of convenient, lysate-free, high throughput tests that can monitor protein phosphorylation and expression profile in cells.

Qualitative determination of target protein concentration is achieved via indirect ELISA.

Key Cell-Based Phospho ELISA Kits


Features

  • Easy-to-use - Conventional methods used to quantify phosphorylated proteins (e.g Western Blot) can be tricky to perform. ELISA Genie's Phospho ELISA kits are much simpler to use and the protocol is less complicated.

  • Convenient - Allow for the simultaneous screening of cell lysate samples at once, without the need to perform Western Blot analysis.

  • Excellent Sensitivity, Specificity, and Efficiency - ELISA Genie Phospho ELISA kits allow for the high throughput screening of several samples with a high signal to noise ratio.

  • Minimal manual labour - No lysate preparation or lysate transfer steps required.

  • Adaptable - Allows for the measurement of target analytes in situ in both adherent and suspension cells.

Contents

Fluorometric Cell-Based Phospho ELISA Kit

  • TBS
  • Quenching Buffer
  • Blocking Buffer
  • Wash Buffer
  • Primary Antibody Diluent
  • α-Phospho-Target Antibody
  • α-Target Antibody
  • α-GAPDH antibody
  • Dye-1 Conjugated Anti-Rabbit IgG Antibody
  • Dye-2 Conjugated Anti-Mouse IgG Antibody
  • 96-Well Black Cell Culture Clear-Bottom Microplate
  • Adhesive Plate Seals

Colorimetric Cell-Based Phospho ELISA Kit

  • TBS
  • Quenching Buffer
  • Blocking Buffer
  • Wash Buffer
  • Primary Antibody Diluent
  • α-Phospho-Target Antibody
  • α-Target Antibody
  • HRP-Conjugated Anti-Rabbit IgG Antibody
  • HRP-Conjugated Anti-Mouse IgG Antibody
  • Crystal Violet Solution
  • SDS Solution
  • Ready-to-Use Substrate
  • Stop Solution
  • 96-Well Cell Culture Clear-Bottom Microplate
  • Adhesive Plate Seals

Applications

  • Quantification of phosphorylated proteins in whole cell cultures

  • Investigation of cell signaling pathways

  • Investigation of pathway activation of other cellular processes (e.g. cell cycle)

Kit Principle

Fluorometric Phospho ELISA

For Fluorometric Cell-Based Phospho ELISA, the target phosphorylated protein is captured by phospho-specific primary (1°) antibodies while Dye 1-conjugated and Dye 2-conjugated secondary (2°) antibodies bind the Fc region of the 1° antibody. Through this binding, the dye conjugated to the 2° antibody can emit light at a certain wavelength given proper excitation, hence allowing for a fluorometric detection method.

A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target RFU values. If a phosphorylated target is being detected, an antibody against the nonphosphorylated counterpart will be provided for normalization purposes.

Figure : Assay format of ELISA Genie Fluorometric Cell-Based ELISA Kit

Colorimetric Phospho ELISA

For Colorimetric Cell-Based Phospho ELISA, the target phosphorylated protein is captured by phospho-specific primary (1°) antibodies while the HRP-conjugated secondary (2°) antibodies bind the Fc region of the 1° antibody. Through this binding, the HRP enzyme conjugated to the 2° antibody can catalyze a colorimetric reaction upon substrate addition.

 


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