Near-infrared Westerns

Introduction

The Azure Imaging Systems has laser technology that offers two near-infrared (NIR) detection channels making it possible to study more than one protein in an assay, even if those targets overlap in molecular weight.

Easily resolve and quantify co-migrating bands, such as phosphorylated and nonphosphorylated isoforms of a protein, or isoforms containing other post-translational modifications. Fluorescent detection provides a larger linear dynamic range than chemiluminescent detection for more accurate quantitation. Imaging with NIR dyes also offers signal stability. An advantage of NIR fluorescent detection over visible fluorescent detection is minimal background because materials including tissue culture plastics demonstrate less autofluorescence in the NIR spectrum.

Fluorescent detection provides a larger linear dynamic range than chemiluminescent detection for more accurate quantitation. An advantage of NIR fluorescent detection over visible fluorescent detection is minimal background because materials including tissue culture plastics demonstrate less autofluorescence in the NIR spectrum. Imaging with NIR dyes also offers signal stability.

Fluorescent Western Blot of STAT1 and phospho-STAT1. The blot was probed with anti-phospho-STAT1 and anti-STAT1 followed by fluorescent secondary antibodies, and then imaged on Azure Imaging system. Top right is the green channel, using IR-800; top left is the image of the red channel, using IR-700. Bottom image is both channels merged.

NIR detection is available on the Azure 500 Imager, as well as the Azure Sapphire Biomolecular Imager.

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Documents

Document TypeDescription
Application NoteMeasuring Histamine Levels in Food SamplesDOWNLOAD
Application NotePerforming an ELISA with the Ao Absorbance Microplate ReaderDOWNLOAD

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