Quantitative Immunofluorescent Assays using Infrared Fluorescence

Application Overview

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Figure 1. Analysis of Akt phosphorylation in MCF-1 breast cancer cells. A) In-Cell Western™ plate image. B) pAkt intensity in 800 channel was normalized to total Akt intensity in 700 channel.

In-Cell Western™ Assay: A rapid, reproducible alternative to traditional Western blotting and ELISA

The In-Cell Western (ICW) Assay is a quantitative immunofluorescence assay performed in microplates (96- or 384-well format). It combines the specificity of Western blotting with the reproducibility and throughput of ELISA. In-Cell Western™ assays are also called cytoblots, cell-based ELISA, In-Cell ELISA (ICE), and FACE (Fast Activated Cell-based ELISA). With In-Cell Western™ assays you can:

  • Detect proteins in fixed cultured cells using target-specific primary antibodies and IRDye® fluorescent secondary antibodies
  • Quantify two targets at 700 nm and 800 nm, using spectrally-distinct infrared dyes
  • Quickly, accurately measure relative protein levels in many samples
  • Detect proteins in situ in a relevant cellular context

ICW assays are used for analysis of:



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