This Application Note describes a new protocol for detecting cell surface protein targets with the Agilent 2100 bioanalyzer and the cell fluorescence LabChip® kit. The specific advantage of an on-chip staining reaction is the amount of time, cells and reagents saved. CD3 on-chip staining on Jurkat cells as well as CD80 (B7-1) and CD86 (B7-2) on- chip staining on 293 cells stably transfected with a CD80 or CD86 expression plasmid were performed. The histogram quality and the percentage of stained cells counted with the microfluidic system are in good agreement with data obtained using a conventional flow cytome- ter. Detailed protocols and reagent recommendations for on-chip stain- ing of cell surface proteins are provided. The new procedure requires 17 fold less cells per sample analysis compared to the conventional staining procedure, while reducing antibody reagent costs by a factor of 80. The high reproducibility of the chip results, very low cell and reagent consumption as well as speed and ease-of-use are advantages the Agilent 2100 bioanalyzer offers for monitoring cell surface protein targets by on-chip antibody staining.
This document details a rapid method to detect protein expression of B7-1 (CD80) and B7-2 (CD86) by performing on-chip antibody staining using the Agilent 2100 bioanalyzer. The procedure utilizes the cell fluorescence LabChip kit to analyze cells in a microfluidic system, offering significant advantages over conventional flow cytometry.
The new procedure requires 17 fold less cells per sample analysis and reduces antibody reagent costs by a factor of 80.
Allophycocyanine (APC) or Cy5 can be used as a dye because the system has high sensitivity in the red with an excitation wavelength of 635 nm.