Reagents available for pilot experiments.

To encourage use of the FACSCalibur and Vantage for multicolor applications, the HSS Flow Core has made available small quantities of PerCP, Red613, Cy5, APC and AMCA-conjugated antibodies and labeling reagents. Talk to Bill if you are interested in trying any of these reagents. They are intended only for pilot experiments.

PerCP. A 488 nm-excited, far red flurochrome useful for four color experiments with FITC, PE and APC or Cy5 on the FACSCalibur. PerCP does not interfere appreciably with APC emission, as do PE-Cy5 tandem conjugates. Click here for examples of four-color analysis with PerCP. We have available:

Red613. A 488 nm-excited, red flurochrome, Red613 is a PE-Texas Red tandem conjugate. It can be used in place of PerCP on the FACSCalibur simultaneous with APC, but requires more compensation. Red613-conjugated reagents are brighter than PerCP, however, making them more useful in some cases. They are also useful for five-color experiments on the Vantage.
APC. A 630 nm-excited, red fluorochrome, APC can be excited both by the diode laser on the FACSCalibur and the helium-neon laser on the Vantage. It can be combined with FITC, PE and PerCP on the FACSCalibur for four-color analysis, or with FITC, PE, Red613 and AMCA for five-color analysis on the Vantage. It is one of the brightest fluorochromes per unit molecule currently characterized. Click here for more detailed info and sample data.
Cy5. A 630 nm-excited, red fluorochrome that can be used interchangeably with APC for four- and five-color experiments. Cy5 is dimmer than APC but its lower molecular weight make it useful for intracellular labeling. Its emission strength is comparable to FITC. Click here for more detailed info and sample data.
AMCA. A 350 nm (UV)-excited, blue fluorochrome, AMCA can be used simultaneously with FITC, PE, Red613 and APC for five-color analysis on the Vantage. Although dimmer than most of the above fluorochromes, it can be conjugated to antibodies against strongly expressed antigens, or used in secondary antibody or biotin-avidin labeling systems for signal enhancement. We have successfully used AMCA to detect b220, Fas and CD8 on mouse lymphocytes. Click here for for info and sample data.
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