Human/Non-Human Primate Regulatory T Cell Staining Kit #1

Also known as: Treg, Foxp3 PCH101; CD4, APC CD25 FITC

RUO: For Research Use Only. Not for use in diagnostic procedures.

SKU# 88-4999

Cat. No.	Size
88-4999-40	Kit

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Description

Description: Regulatory T cells/Tregs (also known as suppressor T cells) are a specialized subpopulation of T cells that act to suppress immune responses thereby maintaining homeostasis and self tolerance. Intense investigation has tried to determine exclusive protein markers for Tregs. This has been complicated by the identification of four regulatory T cells subsets; natural, anergic, Tr1 and CD8+CD28- suppressor T cells. There are still no cell surface molecules that uniquely distinguish functional regulatory cells from conventional T cells. It has been proposed that the “natural” regulatory T cells originally recognized by their constitutive expression of CD4 and CD25 are further defined by expression of the transcription factor Foxp3. Additionally Foxp3 has also been characterized in CD8+CD28- suppressor T cells.

The Complete Regulatory T Cell Staining Kit contains everything needed to stain the natural Tregs including CD4, CD25 and Foxp3 from human, rhesus, chimpanzee or cynomolgus PBMCs. In baboon, CD25 and Foxp3 can be stained, while CD4 cannot be stained using this kit. Other primate species have not been tested.

This kit has been optimized for staining normal hPBMCs. It is critical to use freshly isolated PBMCs to assure optimal Foxp3 staining.

Not included:
Isotype controls for anti-CD4 (mouse IgG1, cat. 11-4714) and anti-CD25 (mouse IgG1, cat. 17-4714)

Details

Reactivity	Human, Monkey
Components	FITC anti-human CD4 (OKT4): 25 tests. Store at 2-8°C in the dark. APC anti-human CD25 (BC96): 25 tests. Store at 2-8°C in the dark. PE anti-human Foxp3 (PCH101): 25 tests. Store at 2-8°C in the dark. PE Rat IgG2a isotype control: 25 tests. Store at 2-8°C in the dark. Normal Rat Serum: 100 µl. Store at 2-8°C. eBioscience Flow Cytometry Staining Buffer: 200 ml. Store at 2-8°C. eBioscience Fixation/Permeabilization Concentrate: 30 ml. Store at 2-8°C. Avoid agitation. This is a 4X stock solution that must be diluted prior to use with the Fixation/Permeabilization Diluent. Dilute 1 part Fixation/Permeabilization Concentrate with 3 parts Fixation/Permeabilization Diluent. Use within 6 months of receipt. Caution: This solution contains Paraformaldehyde, which is toxic and a suspected carcinogen. Contact with eyes, skin and mucous membranes should be avoided. Wear proper protective clothing and gloves. eBioscience Fixation/Permeabilization Diluent: 100 ml. Store at 2-8°C.. The diluent is intended to be used in combination with the Fixation/Permeabilization Concentrate. eBioscience Permeabilization Buffer (10X): 100 ml. Store at 2-8°C. Dilute to 1X with deionized/distilled water and store at 4°C. Caution: Harmful if swallowed or irritant by contact. Wear proper protective clothing and gloves. 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.
Reported Applications	Intracellular Staining Followed by Flow Cytometric Analysis

Documentation

For complete product information, please download the TDS or IFU document.

TDS Link	Download TDS

Additional Formats

References

References: Ahmadzadeh M, Rosenberg SA. IL-2 Administration Increases CD4+CD25hiFoxp3+ Regulatory T Cells in Cancer Patients.Blood. 2005 Nov 22; [Epub ahead of print] (PCH101, intracellular flow, PubMed)

Crellin NK, Garcia RV, Hadisfar O, Allan SE, Steiner TS, Levings MK. 2005. Human CD4+ T Cells Express TLR5 and Its Ligand Flagellin Enhances the Suppressive Capacity and Expression of FOXP3 in CD4+CD25+ T Regulatory Cells. J Immunol. 2005 Dec 15;175(12):8051-9. (PCH101, intracellular flow, PubMed)

Hartwig UF, Nonn M, Khan S, Meyer RG, Huber C, Herr W. Depletion of alloreactive T cells via CD69: implications on antiviral, antileukemic and immunoregulatory T lymphocytes. Bone Marrow Transplant. 2005 Dec 5; [Epub ahead of print] (Intracellular staining for flow cytometry using PCH101, PubMed)

Lim, H.W., P. Hillsamer, A.H. Banham, and C.H. Kim. 2005. Cutting Edge: Direct Suppression of B cells by CD4+CD25+ Regulatory T cells. J. Immunol. 175: 4180-4183. (PCH101, intracellular flow, PubMed)

Manigold T, Shin EC, Mizukoshi E, Mihalik K, Murthy KK, Rice CM, Piccirillo CA, Rehermann B. Foxp3+CD4+CD25+ T cells control virus-specific memory T cells in chimpanzees recovered from Hepatitis C. Blood. 2006 Feb 14; [Epub ahead of print] (PCH101 in chimpanzee, PubMed)

Hori S, Nomura T, Sakaguchi S. 2003. Control of regulatory T cell development by the transcription factor Foxp3. Science. 299(5609):1057-61.

Manavalan JS, Kim-Schulze S, Scotto L, Naiyer AJ, Vlad G, Colombo PC, Marboe C, Mancini D, Cortesini R, Suciu-Foca N.2004. Alloantigen specific CD8+CD28- FOXP3+ T suppressor cells induce ILT3+ ILT4+ tolerogenic endothelial cells, inhibiting alloreactivity. Int Immunol. 16(8):1055-68.

Takahata Y, Nomura A, Takada H, Ohga S, Furuno K, Hikino S, Nakayama H, Sakaguchi S, Hara T. 2004. CD25+CD4+ T cells in human cord blood: an immunoregulatory subset with naive phenotype and specific expression of forkhead box p3 (Foxp3) gene. Exp Hematol. 32(7):622-9.

Sakaguchi S. 2003. The origin of FOXP3-expressing CD4+ regulatory T cells: thymus or periphery.J Clin Invest. 112(9):1310-2.

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