MHC I Streptamers (MHC I T cells) - Cell staining

Staining of antigen-specific MHC I T cells with reversible MHC I Streptamers

The MHC I Streptamers, adducts of Strep-tagged MHC I molecules (MHC I-Streps) and Strep-Tactin¢ç, are directed to antigen-specific CD8 T cell receptors. 
 

Cell staining procedure

1)    Peptide (antigen)-loaded MHC I-Streps (Strep-tagged MHC I molecules) are multimerized with Strep-Tactin¢ç attached to a fluorochrome to generate multimers with high binding avidity. These multimers are determined for staining of T cells which are specific for the respective peptide-loaded MHC molecule. (Fig.1).
Fig. 1: MHC I Streptamer¢ç Fluorochrome
 
2)    After FACS analysis or FACS sorting of the target cells, the staining reagents can be completely removed with biotin (Fig.2). Cells are left phenotypically and functionally indistinguishable from untreated cells.
Fig.2: Antigen-specific cytotoxic CD8 T cells are selected by a MHC I Streptamer¢ç. After staining and fluorescence assisted sorting, the Strep-Tactin¢ç complex is removed from the target cells by the addition of biotin under physiological conditions at low temperatures (4¡ÆC). The monomeric low affinity MHC I-Strep molecules are then dissociated completely from the cell surface by two short washing steps. The isolated cells are phenotypically and functionally undistinguishable from non-treated cells and predestined for clinical use.

Fluorescent MHC I Streptamers for antigen-specific staining of (CD8+) T cells with reversible reagents

streptactinPE.png
 
Fluorescent MHC I Streptamers comprise a MHC I-Strep which has to be associated with a Strep-Tactin¢ç fluorochrome in a short incubation step. The resulting Streptamers are then used for antigen-specific staining of the target cells. After staining and FACS sorting, the Streptamers can be completely removed from the cells with biotin (stock solution included in the Streptamer¢ç Solution Set Standard for washing and dissociation).
 
 
Modular Streptamer¢ç technology : the MHC I-Streps can be associated either with a Strep-Tactin¢ç fluorochrome for cell staining or with a Strep-Tactin¢ç Magnetic Bead for cell isolation.

single MHC I Streptamer¢ç products for cell staining

 
Component
 Use
 Size
 choose among a great variety
 of different alleles and antigens
for staining of 2.5x10^8 cells or for isolation out of 5x10^8 cells human or 2.5x10^8 cells mouse*
 to be associated with MHC I-Streps
for staining of 2.5x10^8 cells*
 Buffer IS 10x for washing
 Biotin stock solution for dissociation
 1 x 50 ml
 1 x 1 ml
 Buffer IS 10x for washing
 Biotin stock solution for dissociation
 1 x 550 ml
 1 x 5 ml
5) Nylon filter tubes  for removal of cell clumps  10 pieces
 
*For larger amounts please refer to the product list or inquire at info@streptamer.com

Best CELLer Staining Kits

Assemble your Best CELLer Staining Kit in two steps:
  1. choose your desired MHC I-Strep (50 tests*) from a great variety of alleles (human, mouse, simian) and more than 80 antigens
  2. select your preferred fluorochrome (50 tests* of Strep-Tactin PE or Strep-Tactin APC)
Please use filters "Category" and "Subcategory" on top of the  product list
* 1 test is for staining of 1x10^6 cells.
 
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