Choosing Labels and Conjugating Antibodies for Multicolor Flow Cytometry and Other Immunoassays

Target-specific antibodies, conjugated to affinity or fluorogenic small-molecule tags, are powerful tools for quantifying and localizing proteins. Antibody labeling kits are popular, but can be time-consuming and risk sample loss. In this webinar, Dr. Timothy Bushnell (University of Rochester Medical Center) will discuss the state of the art in multicolor panel design, and address important considerations and inherent challenges. Dr. Amedeo Cappione III (MilliporeSigma) will then outline an alternative method for small-scale protein labeling based upon continuous flow diafiltration in a centrifugal filter. The method is simple, fast, and has few hands-on requirements, without sacrificing labeling efficiency, final yield, or conjugate performance.

What You Will Learn:
- How to overcome the challenges of multicolor panel design
- Best practices for optimizing the ratio between fluorophore and antibody
- The importance of considering the buffer components in which commercial antibodies are provided
- How to use centrifugal diafiltration for fast and effective buffer exchange and concentration of antibody solutions, even when amount of antibody is very limited.

Speakers:
Timothy Bushnell, PhD
Research Director
University of Rochester Medical Center (URMC)

Amedeo Cappione III, PhD
Senior Scientist
MilliporeSigma

Original Broadcast: June 26th, 2016