Human/Mouse/Rat FoxP3 Alexa Fluor® 647-conjugated Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human FoxP3 in direct ELISAs. Detects human, mouse, and rat FoxP3 in flow cytometry.
  • Source
    Recombinant Monoclonal Mouse IgG1 Clone # 376209
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    E. coli-derived recombinant human FoxP3
    Gln105-Lys200
    Accession # Q9BZS1
  • Formulation
    Supplied in a saline solution containing BSA and Sodium Azide.
  • Label
    Alexa Fluor 647
Applications
  •  
    Recommended
    Concentration
    Sample
  • Intracellular Staining by Flow Cytometry
    5 µL/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Intracellular Staining by Flow Cytometry
Detection of FoxP3 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human/Mouse/Rat FoxP3 Alexa Fluor® 647‑conjugated Monoclonal Antibody (Catalog # IC8970R) and Mouse Anti-Human CD4 Fluorescein‑conjugated Monoclonal Antibody (Catalog # FAB3791F). Quadrant markers were set based on control antibody staining (Catalog # IC002R). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Intracellular Staining by Flow Cytometry
Detection of FoxP3 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human/Mouse/Rat FoxP3 Alexa Fluor® 647‑conjugated Monoclonal Antibody (Catalog # IC8970R) and Mouse Anti-Human CD25/IL-2 R alpha PE‑conjugated Monoclonal Antibody (Catalog # FAB1020P). Quadrant markers were set based on control antibody staining (Catalog # IC002R). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Intracellular Staining by Flow Cytometry
Detection of FoxP3 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Mouse Anti-Human/Mouse/Rat FoxP3 Alexa Fluor® 647‑conjugated Monoclonal Antibody (Catalog # IC8970R) and Rat Anti-Mouse CD4 Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # FAB554G). Quadrant markers were set based on control antibody staining (Catalog # IC002R). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Intracellular Staining by Flow Cytometry
Detection of FoxP3 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Mouse Anti-Human/Mouse/Rat FoxP3 Alexa Fluor® 647‑conjugated Monoclonal Antibody (Catalog # IC8970R) and Rat Anti-Mouse CD25/IL-2 R alpha PE‑conjugated Monoclonal Antibody (Catalog # FAB2438P). Quadrant markers were set based on control antibody staining (Catalog # IC002R). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Intracellular Staining by Flow Cytometry
Detection of FoxP3 in Rat Splenocytes by Flow Cytometry. Rat splenocytes were stained with Mouse Anti-Human/Mouse/Rat FoxP3 Alexa Fluor® 647‑conjugated Monoclonal Antibody (Catalog # IC8970R) and Anti-Rat CD4 APC-conjugated Antibody. Quadrant markers were set based on control antibody staining (Catalog # IC002R). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Protect from light. Do not freeze.
    • 12 months from date of receipt, 2 to 8 °C as supplied.
Background: FoxP3
Human FoxP3 is a 431 amino acid (aa), 47 kDa member of the P subclass of the FOX (forkhead box) family of transcription factors. It contains a Leu-rich repeat, a C2H2 zinc finger region, and a C-terminal FKH (fork head), DNA-binding domain. Three alternative splicing events have been described in human. The first shows a deletion of aa 72-106, a second exhibits a deletion of aa 246-272, and a third records a 60 aa insertion after Lys382. These events are likely not to be mutually exclusive. The first isoform (plus the full-length form) occurs in CD4+CD25+ T cells, while the less frequent second isoform appears in CD8+ Tregs. The mouse FoxP3 gene does not appear to be spliced. In addition, FoxP3 is found only in Tregs in mouse, while human FoxP3 appears in both Tregs and CD4+ and CD8+ conventional T cells.

Both mouse and human FoxP3 undergo proteolytic processing, after Arg51 and Arg417 in human, and at equivalent sites in mouse. Cleavage at the N-terminus generate a 41 kDa isoform, whereas C-terminal cleavage generates an isoform indistinguishable from the full-length isoform (47 kDa) in SDS-PAGE.

FoxP3 can act as a transcriptional repressor (for IL-2) via an interaction with HDAC7. By contrast, it can also act as a transcriptional activator (for GITRL and
CTLA-4) through an interaction with RUNX7. Notably, for both human and mouse, full-length 47 kDa FoxP3 will induce high levels of CTLA-4 on expressing cells. However, when the C-terminally truncated isoform (at Arg417) is considered, IL-10 promoting activity is enhanced in mouse, while suppressive activity is lost in human. Over aa 105-200, human FoxP3 shares 83% and 84% aa sequence identity with rat and mouse FoxP3, respectively.
  • Long Name:
    Forkhead Box P3
  • Entrez Gene IDs:
    50943 (Human); 20371 (Mouse)
  • Alternate Names:
    AIID; AIIDMGC141961; DIETER; forkhead box P3; Forkhead Box Protein P3; FoxP3; FOXP3delta7; immune dysregulation, polyendocrinopathy, enteropathy, X-linked; Immunodeficiency, Polyendocrinopathy, Enteropathy, X-Linked; IPEX; JM2; MGC141961; MGC141963; PIDX; PIDXMGC141963; SCURFIN; XPID; XPIDpolyendocrinopathy, enteropathy, X-linked
Related Research Areas
Isotype Controls
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Mouse IgG1 Alexa Fluor® 647-conjugated Antibody

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