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6-8000-2011 Kit$699.00   $799.00Buy Now | Add to Cart
 
Type: Cell Isolation-Separation
 
Applications: Cell Assays
E=ELISA; FACS; FC=Flow Cytometry; FPLC=Fast Protein Liquid Chromatography; GF=Gravity Flow; HPLC=High Performance Liquid Chromatography; ICC=Immunocytochemistry; IF=Immunofluorescence; IHC=Immunohistochemistry; IP=Immunoprecipitation; NAC=Non-adherent Cell Assays; NB=Neutralization of Bioactivity; SE=Sandwich ELISA; TPE=Targeted Protein Expression; WB=Western blotting; ; AC=Adherent Cell Assays; FM=Fluorescent Micsroscopy; ; ; BSC-CM5= Biacore Sensor Chip CM5; BSM=Biosactive Small Molecule or Peptide; CDM=Cell Differentiation Media; ; ; ; ; ; Health and Fitness; ; ; DNA Extraction/Purification; ; In vivo Like Assays
Species Reactivity: H
B=Bovine; Ca=Cat; Ch=Chicken; D=Dog; EQ=Equine; GP=Guinea Pig; H=Human; M=Mouse; P=Porcine; Pr=Primate; R=Rat; Rb=Rabbit; Y=Yeast; Xe=Xenopus; Ze=Zebrafish; ; ; ; NA-Not Applicable; STP=Step-Tactin Proteins; All
Description/Data:
Data for CD3 Fab Streptamer Isolation

For isolation of CD3+ T cells from peripheral blood mononuclear cells. The kit includes all Fab Streptamer components for positive cell isolation with and for removal of Streptamers from isolated cells:

  • Strep-Tactin® Magnetic Microbeads* for Fab Streptamers: 2 vials for 5x108 cells each
  • CD3 Fab-Strep human for isolation of T-cells: 1 vial for 1x109 cells
  • D-Biotin stock solution for Streptamer® technology: 2x 1 ml
  • Buffer IS, Streptamer® Washing Buffer, 10x concentrated: 50 ml

Figure: Streptamer® CD3/CD28 premix for T cell expansion. Human T cells isolated with the CD3 Fab Streptamer® Isolation Kit MB (68000-201) were cultured for 13 days with the CD3/CD28 Streptamers® for cell expansion. Fold expansion was measured.

*Microbeads are intended for use with a permanent magnet for manual cell isolation; we recommend our StrepMan magnet for 2 x 15 ml and 2 x 50 ml Falcon tubes. Note: Kit price includes dry ice and handling charges.

Isolation of human CD3+ T cells is performed by positive selection. PBMCs are labeled with magnetic CD3 Fab Streptamers. Labeled cells are isolated in a strong magnet (StrepMan, horizontal position) where they migrate toward the tube wall on the side of the magnet. This CD3 positive cell fraction is collected and cells are liberated from all labeling reagents by addition of biotin in a strong magnet (StrepMan, upright position). The magnetic Streptamers migrate toward the tube wall and the label-free cells remain in the supernatant. Biotin is removed by washing.

The resulting cell preparation is highly enriched with CD3+ T cells with a purity of more than 90%. No depletion steps and no columns are needed.