Foxp3 Mouse Monoclonal Antibody [Clone ID: 3G3]

CAT#: AM12131PU-N

Foxp3 mouse monoclonal antibody, clone 3G3, Aff - Purified

Product Images

Intracellular staining of human peripheral blood cells (gated on CD4+ cells) with anti-FoxP3 (clone 3G3).

Specifications

Product Data

• Clone Name: 3G3
• Applications: FC, WB
• Recommended Dilution: Flow cytometry (Application note see below).
  Western blot: 2 µg/ml.
• Reactivities: Human, Mouse
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Full-length His-tagged recombinant murine FoxP3
• Specificity: The mouse monoclonal antibody 3G3 recognizes N-terminal region of FoxP3, a 47-55 kDa transcription factor, which is the master regulator in the development and function of regulatory T cells.
• Formulation: Phosphate buffered saline (PBS) with 15 mM sodium azide, approx. pH 7.4
  State: Aff - Purified
  State: Liquid Ig fraction
• Concentration: 1.0 mg/ml
• Purification: Protein A affinity chromatography
• Gene Name: Mus musculus forkhead box P3 (Foxp3), transcript variant 2
• Database Link:
  Entrez Gene 50943 HumanEntrez Gene 20371 Mouse
• Background: FoxP3 (Forkhead box protein 3), a highly conserved forkhead/winged-helix transcription factor, plays a crucial role in maintaining immune homeostasis by governing the development and function of regulatory T cells. It is constitutively expressed at high level in CD25+ CD4+ Treg cells and at low level in a CD25- CD4+ Treg cell subset. Defects in gene encoding FoxP3 protein cause the scurfy phenotype in mice, and in human the IPEX syndrome (immune dysfunction, polyendocrinopathy, enteropathy, X-linked syndrome), also known as X-linked autoimmunity-allergic dysregulation (XLAAD) syndrome.
• Synonyms: Forkhead box protein P3, IPEX, JM2, Scurfin
• Note: Application note for Flow cytometry:
  Staining method:
  - Perform staining of cell surface markers (CD25, CD4 etc.) for 20 min. in the dark and RT. Wash.
  - Add cold fixation buffer diluted to working concentration (if commercial reagent is used, follow the manufacturer’s instructions) and incubate for 30-45 min. in 4°C.
  - Centrifugate (200 g, 5 min., 4°C) and add a cold permeabilization buffer diluted to working concentration (if commercial reagent is used, follow the manufacturer’s instructions) to the pellet of cells.
  - Centrifugate, and add to the pellet 100 µl of the permeabilization buffer and 5-50µl of blocking solution (NMS, FTS, 1% BSA etc., or commercial blocking solutions) incubate for 15 min. (4°C, in the dark).
  - Perform intracellular staining of FoxP3 for 30 min. (4°C, in the dark) with appropriate amount of 3G3 antibody (final concentration 2-10 µg/ml; optimal concentration for peripheral blood is 3 µg/ml).
  - Centrifugate (200 g, 5 min., 4°C) and add 4 ml of cold permeabilization buffer to the pellet.
  - Centrifugate again (200 g, 5 min., 4°C) and add 100 µl of staining buffer (PBS with 0.2% BSA). Keep cold until measuring on a flow cytometry device with appropriate setting.
  (If necessary, for the same sample use mouse IgG1 isotype control in the same format as anti-FoxP3 antibody 3G3.)