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Detection of antibody-stained cell,,,surface and intracellular protein targets,,,with the Agilent 2100 bioanalyzer

This Application Note describes how the Agilent 2100 bioanalyzer and the Cell Fluorescence LabChip kit can be used to detect cell surface and intracellular protein targets by antibody staining. CD3 staining on Jurkat cells as well as CD4 staining on CCRF-CEM cells and intracellular glucocorticoid receptor (GR) staining of H4 rat hepatocytes were performed. Histogram quality and the percentage of stained cells counted with the microfluidic system are in good agreement with data obtained with a conventional flow cytometer. Detailed protocols and reagent recommendations for staining cell surface and intracellular protein targets are given. The high reproducibility of the chip results, low cell consumption and ease-of-use are advantages of the compact bioanalyzer system for monitoring cell surface and intracellular protein target expression by antibody staining.
The Agilent 2100 bioanalyzer was introduced by Agilent Technologies as the first commercially available lab-on-a-chip analysis system for the life science laboratory using LabChip products developed by Caliper Technologies Corp. Chip-based approaches for a variety of separation-based techniques have been introduced, addressing DNA, RNA, and protein separations. The Agilent 2100 bioanalyzer is capable of two-color fluorescence detection and runs disposable microfluidic glass chips. The applications presented here are based on the controlled movement of cells by pressure-driven flow inside the interconnected networks of microfluidic channels. Cells are hydrodynamically focused in these channels before passing the fluorescence detector in single file. Each chip accommodates up to six samples and data acquisition of all samples is fully automated while data analysis allows for user-specific settings.
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