Foxp3 Mouse Monoclonal Antibody [Clone ID: 3G3]

Specifications

Product Data
Clone Name	3G3
Applications	FC, WB
Recommend Dilution	Flow cytometry (Application note see below). Western blot: 2 µg/ml.
Reactivity	Human, Mouse
Host	Mouse
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	lot specific
Purification	Protein A affinity chromatography
Conjugation	Unconjugated
Storage Condition	Store at 2 - 8 °C for up to one month or (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Gene Name	forkhead box P3
Database Link	Entrez Gene 50943 HumanEntrez Gene 20371 Mouse UniProt ID Q99JB6
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.)
Reference Data

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