Application data MHC I Streptamer® for isolation of CD8+ T cells

Staining of antigen-specific CD8+ T cells with reversible MHC I Streptamer®

High staining intensities with MHC I Streptamer®: High staining intensity with Streptamer® guarantees optimal detection of small T cell populations. Ex vivo staining of CMV-specific CD8+ T cells with HLA-A*0201 CMV pp65495-503 Streptamer® (right diagram) or conventional multimers (left diagram). Staining was performed from a CMV positive individual at 4°C.

 

Used Streptamer® reagents:
  • 6-7001-001 MHC I-Strep CMV, pp65495-503
  • 6-5000-001 Strep-Tactin® PE
  • 6-5603-005 Streptamer® Solution Set Standard for washing and dissociation

Staining and removal of staining from antigen-specific mouse CD8+ T cells

Staining of bacteria-specifc (Listeria monocytogenes, epitope LLO91-99) CD8+ T-cells with H2-Kd/LLO91-99 Streptamers®. Antigen-specific cells were visualized with Strep-Tactin® PE coupled to MHC I-Strep H2-Kd specific for LLO91-99 . After addition of 1 mM d-biotin, subsequent dissociation of monomerized MHC molecules was monitored at different time points as indicated. 20 minutes after biotin addition, staining reagents were largely removed from the cells. Staining and dissociation were performed at 4 °C.

Used Streptamer® reagents:
  • 6-7014-001 MHC I-Strep H2-Kd, LLO91-99
  • 6-5000-001 Strep-Tactin® PE
  • 6-5603-005 Streptamer® Solution Set Standard for washing and dissociation

Staining and removal of staining from antigen-specific human CD8+ T cells

Staining of Her2/neu-specifc CD8+ T cells (kindly provided by H. Bernhard, TU Munich) with HLA-A*0201/Her2/neu369-377 Streptamers® as compared to conventional multimers (left diagram). Complete dissociation of Streptamer® staining reagents from the cells is shown after a brief incubation (30 min.) in the presence of 1 mM d-biotin (right diagram). Antigen-specific T cells were visualized with Strep-Tactin® PE coupled to MHC I-Strep HLA-A*0201 specific for Her2/neu369-377. Staining and dissociation were performed at 4 °C.

Used Streptamer® reagents:
  • 6-7008-001 MHC I-Strep HLA-A*0201, Her2/neu369-377
  • 6-5000-001 Strep-Tactin® PE
  • 6-5603-005 Streptamer® Solution Set Standard for washing and dissociation

All experiments were performed in close cooperation with D. Busch, TU Munich.