Skip to main content
. Author manuscript; available in PMC: 2010 Oct 13.
Published in final edited form as: J Immunol Methods. 2009 Jan 31;343(1):28–41. doi: 10.1016/j.jim.2009.01.005

Fig. 6. Identifying antigen-specific hybridomas by FACS.

Fig. 6

Examples of FACS analysis performed on four antigens prior to single cell deposition of potential hybridomas into microtiter plates. Hybridomas were sorted first (vertical axis) based on reactivity with various antigens: A, Influenza A/Jb (Johannesburg); B, KLH; C, ARP5; D, HSV2gG glycoprotein; and second (horizontal axis) based on reactivity with GAM Ig. Cells were labeled with biotin-conjugated antigen coupled with PE-conjugated streptavidin and APC-conjugated GAM Ig (H+L). Squares highlight sorted populations: Q1 (orange, labeled for antigen only), Q2 (yellow, labeled for antigen and surface Ab), Q3 (blue, unlabeled), and Q4 (red, labeled for surface Ab only). Only 10,000 events are shown on each plot for simplicity of interpretation. The actual number events per sorting experiment generally exceeded 3×107 (roughly 3 hours sorting at 3000 events/second).