Total Protein Normalization

While fluorescent and chemiluminescent imaging techniques and their detectors are becoming more sensitive and selective, photostimulated luminescence from phosphor storage plates remains a key application for many researchers, especially for applications such as electrophoretic mobility shift assays (EMSA), enzyme assays, and in-vivo imaging.

Traditional phosphor imagers are often single function and require a large footprint. The Azure
Sapphire Biomolecular Imager offers the ability to scan storage phosphor screens for filmless autoradiography, with exceptional dynamic range and image quality thanks to the use of laser excitation and photon multiplier tube (PMT) detection. The Sapphire also incorporates exceptional visible fluorescent and near infra-red scanning along with true chemiluminescent and white light imaging making it a true lab workhorse.

Materials and Methods
Sample Preparation

An American Radiolabeled Chemicals 14C standard with slices from 1000 to 0.004 μCi/g was exposed to a BAS-MS storage phosphor screen (0.9 DPM/mm2/hr. sensitivity) for 3 hours.

Storage Phosphor Plate Imaging

Following exposure, the storage phosphor screen was imaged using the Phosphor Imaging module of the Sapphire with standard excitation and detection settings. Samples were quantified and limits of detection (LOD) and dynamic range (DR) calculated.

Results and Conclusions

The Sapphire captures high quality images with a large dynamic range from standard storage phosphor screens (Figure 1). Following sample quantification using AzureSpot analysis software, the LOD for American Radiolabeled Chemicals 14C standard after a 3-hour exposure was determined to be 0.036 μCi/g. Furthermore, a highly linear dynamic range from 1000 to 0.036 μCi/g was observed (r2 = 0.99 [Figure 2]).

To conclude, the Sapphire is capable of imaging storage phosphor screens producing high quality images for later quantification, while also offering the ability to rapidly image fluorescent, chemiluminescent and NIR samples all within a single compact footprint.

Materials and Product Numbers
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Application NoteTotal Protein Normalization with AzureRedDOWNLOAD
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