Invitrogen™ eBioscience™ Human Regulatory T Cell Staining Kit #2

Invitrogen™ eBioscience™ Human Regulatory T Cell Staining Kit #2

This Human Regulatory T cell Staining Kit contains all of the buffers and monoclonal antibodies for CD4, CD25, and Foxp3 necessary to successfully stain and identify regulatory T cells from human peripheral blood cells.

The RPA-T4 monoclonal antibody reacts with human CD4, a 59 kDa glycoprotein found on the surface of the majority of thymocytes, a subset of mature T cells (T helper cells), and at lower levels on monocytes. The BC96 monoclonal antibody reacts with human CD25 (also known as interleukin-2 receptor alpha, IL-2R alpha), a 55 kDa surface protein expressed by early progenitors of T cells and B cells, by mature, activated T cells and B cells, and at constitutively high levels on regulatory T cells. The PCH101 monoclonal antibody reacts with the amino terminus of human Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2. Foxp3 is a 49-55 kDa protein and a member of the forkhead/winged-helix family of transcription factors. It was identified as the gene responsible for the X-linked lymphoproliferative disease observed in scurfy (sf) mice and in the human disorder, X-linked autoimmunity-allergic dysregulation syndrome (XLAAD). Constitutive expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Content

• Flow Cytometry Staining Buffer (cat. 00-4222): 200 mL, store at 2-8°C.
• Fixation/Permeabilization Concentrate (4X) (cat. 00-5123): 30 mL, store at 2-8°C. Avoid agitation.
• Fixation/Permeabilization Diluent (cat. 00-5223): 100 mL, store at 2-8°C.
• Permeabilization Buffer (10X) (cat. 00-8333): 100 mL, store at 2-8°C. Note: The 10X Permeabilization Buffer has a natural tendency to precipitate, however, its function is not affected by this. To clarify, the solution can be filtered after dilution to 1x working solution.
• Normal Rat Serum (cat. 24-5555): 100 μL, store at 2-8°C.
• eZFluor™ Anti-Human CD4 FITC and CD25 PE Cocktail (clones RPA-T4 and BC96, cat. 22-8425): 25 tests, store at 2-8°C. Light-sensitive material.
• Anti-Human Foxp3 APC (clone PCH101, cat. 17-4776): 25 tests, store at 2-8°C. Light-sensitive material.
• Rat IgG2a K Isotype Control APC (cat. 77-4321): 25 tests, store at 2-8°C. Light-sensitive material.