This product consists of a Foxp3 Transcription Factor Staining Buffer Kit (A24261) and a Foxp3 Mouse anti-Mouse mAb (clone 3G3), PE conjugate (A18690).
About the Antibody
The Foxp3 Mouse Anti-mouse mAb (clone 3G3), PE conjugate, is a monoclonal antibody that reacts with mouse Foxp3 protein. R-PE is a stable and highly soluble phycobiliprotein that provides maximal absorbance and fluorescence emission without susceptibility to internal or external fluorescence quenching, thus providing exceptional quantum yields and molar extinction coefficients. Use of R-PE in antibody conjugates results in probes with greatly enhanced detectability.
About the Buffers
The buffers in the included Foxp3 Transcription Factor Buffer Kit have been specially formulated to work with the PE conjugate of Foxp3 Mouse Anti-Mouse mAb (clone 3G3) for optimal observation of intranuclear staining when used in flow cytometry applications. The Foxp3 Transcription Factor Buffer Kit is compatible with intracellular cytokine staining within the same panel.
The kit contains 3 buffers:
• Foxp3 Fixation Transcription Factor and Permeabilization Buffer A (A24217, a 4X concentrate)
• Foxp3 Transcription Factor Fixation and Permeabilization Buffer B (A24218, a diluent)
• Foxp3 Transcription Factor Wash Buffer (A24261, 10X)
How the Buffers Work
The Foxp3 Transcription Factor Fixation and Permeabilization Buffer A is supplied as a 4X stock solution and must be diluted with Foxp3 Transcription Factor Fixation and Permeabilization Buffer B prior to use.
The Foxp3 Transcription Factor Wash Buffer is supplied as a 10X stock solution. The 10X stock solution should be diluted to a 1X working concentration in PBS containing 1% BSA and used for wash and intranuclear Foxp3 staining steps following fixation and permeabilization with 1X Foxp3 Fixation and Permabilization buffers.
About Foxp3
The 3G3 antibody reacts with mouse Foxp3 protein, a 50-55 kDa transcription factor that is a master regulator of regulatory T cell (Treg) development and function. Foxp3 is expressed constitutively by Tregs, which are further identified as being CD4+ CD25+, whereas resting conventional CD4+ T cells do not express Foxp3. However, TCR stimulation of naive T cells in the presence of TGF-beta has been shown to induce expression of Foxp3, driving their differentiation into Foxp3+ Tregs, which are called “induced" or “adaptive" Tregs. These cells are phenotypically similar to "natural" Tregs (CD4+ CD25+ Foxp3+), which originate in the thymus and comprise the majority of Tregs in the periphery. Tregs are critical for maintaining peripheral immune homeostasis. Defects in Foxp3 expression or Treg function have been implicated in the development of autoimmunity. Tregs have also been shown to restrict immune responses to infection, cancer, and vaccination. The 3G3 antibody may be used for intracellular detection of Foxp3 in cells from mouse and Rhesus macaque.
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