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 document details how the Agilent 2100 bioanalyzer, in combination with the Cell Fluorescence LabChip kit, can be used to detect cell surface and intracellular protein targets through antibody staining. It covers protocols for extracellular CD3 and CD4 staining as well as intracellular glucocorticoid receptor (GR) staining.
The system offers high reproducibility, low cell consumption, and ease-of-use.
APC or Cy5 can be used as dyes because the bioanalyzer has high sensitivity in the red.