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 Firefly™ 3000 Protein Assay System Cell BioSciences FireFly™ 3000 system separates and quantifies proteins in scarce and complex matrices, delivering the most significant improvement to whole cell protein quantitation since the development of Western Blot assays. FireFly combines two orthogonal techniques, physical separation using isoelectric focusing and specific antibody binding using secondary antibody detection, to separate and quantify specific proteins. Using disposable capillaries with a total volume of 400 nanoliters, Firefly enables researchers, for the first time, to measure proteins in scarce and valuable samples such as stem cell, primary cells, and fine needle aspirates.The system uses standard 96-well plates, enabling unattended analysis of dozens of samples. Single Antibody Detection
Single Antibody Detection
Quantitative Analysis of Erk Kinases with Total Erk Antibody
Data courtesy: Richard Neve, UCSF Erk1 and Erk2, which are activated by phosphorylation of a threonine and tyrosine, are key kinases in the MAP cascade that play a key role in cell growth. Singly and doubly phosphorylated forms of Erk1 and Erk2 are separated by isoelectric focusing and then immobilization and detection on-capillary. Immunodection uses antibodies against total Erk1 and Erk2, eliminating the need for phospo-specific antibodies. Note that Western blotting, which separates proteins based on size, does not resolve these different phosphorylated forms.
Three breast cancer lines were compared with a normal tissue line, MFC10A. Note the significant differences in the ratios and levels of mono and dual phosphorylated forms of Erk1 and Erk2 across the breast cancer lines. Small Sample Size
Firefly analyses are routinely performed with less than 1000 cells. The limit of detection is analyte specific, but has been demonstrated at below 25 cells.  Firefly’s modest sample requirements enable use of Fine Needle Aspirates, allowing for serial sampling. Western blots often require Mouse sacrifices to collect sufficient protein samples.
Quantitative and Sensitive
Measurement of proteins is fast and quantitative with Firefly. All separations occur in a short length of capillary, with a total fixed volume of 400 nL, ensuring sample-to-sample uniformity and consistent results. Chemiluminescence over the entire length of capillary is measured with a sensitive CCD camera, providing quantitation Automation
After loading samples and reagents using a standard 96-well plate, the system runs completely unattended. Applications
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