PRODUCT CODE: ET1702-12

Recombinant FOXP3 Monoclonal Antibody (ET1702-12)

  • Recombinant

Applications

  • ICC

  • FC

REACTIVITY

  • Human

Western blot analysis of FOXP3 on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-12, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of FOXP3 on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-12, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of FOXP3 in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-12, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of FOXP3 in 293T cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-12, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Flow cytometric analysis of FOXP3 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-12, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of FOXP3 on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-12, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • ICC

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant FOXP3 Monoclonal Antibody (ET1702-12)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

K562 cell lysates, MCF-7, 293T, Jurkat.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JF0898

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

Isotype

IgG

APPLICATION DILUTION

  • ICC

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

FOXP3

SYNONYMS

AIID antibody; DIETER antibody; Forkhead box P3 antibody; Forkhead box protein P3 antibody; FOXP3 antibody; FOXP3_HUMAN antibody; FOXP3delta7 antibody; Immune dysregulation polyendocrinopathy enteropathy X linked antibody; Immunodeficiency polyendocrinopathy enteropathy X linked antibody; IPEX antibody; JM2 antibody; MGC141961 antibody; MGC141963 antibody; OTTHUMP00000025832 antibody; OTTHUMP00000025833 antibody; OTTHUMP00000226737 antibody; PIDX antibody; Scurfin antibody; XPID antibody

POST-TRANSLATIONAL MODIFICATION

Polyubiquitinated, leading to its proteasomal degradation in regulatory T-cells (Treg) which is mediated by STUB1 in a HSPA1A/B-dependent manner. Deubiquitinated by USP7 leading to increase in protein stability.; Phosphorylation at Ser-418 regulates its transcriptional repressor activity and consequently, regulatory T-cells (Treg) suppressive function. Dephosphorylated at Ser-418 by protein phosphatase 1 (PP1) in Treg cells derived from patients with rheumatoid arthritis. Phosphorylation by CDK2 negatively regulates its transcriptional activity and protein stability (By similarity).; Acetylation on lysine residues stabilizes FOXP3 and promotes differentiation of T-cells into induced regulatory T-cells (iTregs) associated with suppressive functions. Deacetylated by SIRT1.; Undergoes proteolytic cleavage in activated regulatory T-cells (Treg), and can be cleaved at either the N- or C-terminal site, or at both sites.

SUBCELLULAR LOCATION

Nucleus, Cytoplasm.

FUNCTION

The FOX family of transcription factors is a large group of proteins that share a common DNA binding domain termed a winged-helix or forkhead domain. During early development, FOXP1 and FOXP2 are expressed abundantly in the lung, with lower levels of expression in neural, intestinal and cardiovascular tissues, where they act as transcription repressors. FOXP1 is widely expressed in adult tissues, while neoplastic cells often exhibit a dramatic change in expression level or localization of FOXP1. The gene encoding human FOXP1 maps to chromosome 3p14.1, and the gene encoding human FOXP2 maps to chromosome 7q31. The gene encoding FOXP3, a third member of this family, maps to chromosome Xp11.23. Mutations in this gene cause IPEX, a fatal, X-linked inherited disorder characterized by immune dysregulation. The FOXP3 protein, also known as scurfin, is essential for normal immune homeostasis. Specifically, FOXP3 represses transcription through a DNA binding forkhead domain, thereby regulating T cell activation.