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Products:Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
Overview
Product name
Anti-FOXP3 antibody
See all FOXP3 products (28) ...
Description
Rabbit polyclonal to FOXP3
Specificity
This antibody recognises a single clean band in a HEK293 cell lysate that overexpresses human FOXP3. The band is not seen in untransfected cells and is blocked by the immunising peptide. The antibody has not been tested in WB against endogenous FOXP3. All 3 human FOXP3 isoforms will be detected when the antibody is tested against endogenous samples.
Tested applications
IHC-P, Flow Cyt, WB, IHC-Fr, ICC, ICC/IFmore details
Cross reactivity
Reacts with
Human
Predicted to work with
Macaque Monkey
Does not react with
Mouse
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human FOXP3.
(Peptide available as ab14151.)
Positive control
ICC: U2OS cells WB: HEK 293 lysate over expressing human FOXP3 IHC-Fr: human tonsil FACS: purified human CD4+, CD4+CD25- and CD4+CD25+ cells (see review)
General notes
There is a reported difference in the human and mouse immune systems regarding FOXP3 expression being induced by T cell activation.
In contrast to the murine system, the activation of human CD4+ CD25- T cells through TCR leads to the increased expression of FOXP3 mRNA, the surface expression of CD25 and gain of regulatory function (see references especially Walker et al, 2003). This suggests that FOXP3 in human is not a unique marker for regulatory T cells.
The pattern of staining reported with ab10563 is consistent with the pattern of other anti-FOXP3 antibodies commercially available and we would welcome your opinion regarding the difference in population stained with FOXP3 antibodies; certain users have indeed observed a staining in human CD25- cells as reported in several papers.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Immunology >> Adaptive Immunity >> Regulatory T Cells
Epigenetics and Nuclear Signaling >> Transcription >> Domain Families >> Forkhead Box >> FOXP
Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
Applications
Show applications key
Our Abpromise guarantee covers the use of ab10563 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P
Flow Cyt
No star rating (1 Abreview)ICC
Application notes
Flow Cyt: Use at a concentration of 4 µg/ml.
ICC: Use at a concentration of 2.5 - 5 µg/ml.
IHC-P: Use at an assay dependent dilution (PMID 19196820).
IHC-Fr: Use at a concentration of 0.625 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately ~50 kDa (predicted molecular weight: 47.2 kDa). Can be blocked with FOXP3 peptide (ab14151).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
Function
Probable transcription factor. Plays a critical role in the control of immune response.
Involvement in disease
Defects in FOXP3 are the cause of immunodeficiency polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) [MIM:304790]; also known as X-linked autoimmunity-immunodeficiency syndrome. IPEX is characterized by neonatal onset insulin-dependent diabetes mellitus, infections, secretory diarrhea, trombocytopenia, anemia and eczema. It is usually lethal in infancy.
Sequence similarities
Contains 1 C2H2-type zinc finger.
Contains 1 fork-head DNA-binding domain.
Cellular localization
Nucleus.
Target information above from: UniProt accessionQ9BZS1
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- AIID antibody
- DIETER antibody
- Forkhead box P3 antibody
see all
Database links
- Entrez Gene: 50943 Human
- Omim: 300292 Human
- SwissProt: Q9BZS1 Human
- Unigene: 247700 Human
Anti-FOXP3 antibody images:
Immunofluorescence - FOXP3 antibody (ab10563)
Immunofluorescent imaging of human cells (U2OS) with ab10563 (5 ug/ml) confirms the specificity of this anti-FOXP3 antibody, which recognises nuclear signal only; with no background cytoplasmic or nucleolar staining.
IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour). Primary antibody used at 5ug/ml in 5% milk / 0.2% TWEEN for 1 hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees C. Nuclei were stained with Hoechst stain.
Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK
Immunohistochemistry (Frozen sections) - FOXP3 antibody (ab10563)
FoxP3 immunostaining on a frozen section from human tonsil. Original magnification x100. Nuclear staining was observed.
Cigdem Atayar
Western blot - FOXP3 antibody (ab10563)
Lanes 1 - 6 : Anti-FOXP3 antibody (ab10563) at 2 µg/ml
Lane 7 :
Lane 1 : 20µg HEK 293 lysate
Lane 2 : 20µg HEK 293 lysate over expressing human FOXP3
Lane 3 : 20µg HEK 293 lysate over expressing mouse FOXP3
Lane 4 : 20µg HEK 293 lysate with FOXP3 peptide (ab14151) at 1 µg/ml
Lane 5 : 20µg HEK 293 lysate over expressing human FOXP3 with FOXP3 peptide (ab14151) at 1 µg/ml
Lane 6 : 20µg HEK 293 lysate over expressing mouse FOXP3 with FOXP3 peptide (ab14151) at 1 µg/ml
Lane 7 : 20µg HEK 293 lysate
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 0.2 µg/ml
Performed under reducing conditions.
Predicted band size : 47.2 kDa
Observed band size : 47 kDa (why is the actual band size different from the predicted?)
Exposure time : 30 seconds
This antibody recognises a single clean band in a HEK293 cell lysate that overexpresses human FOXP3. The band is not seen in untransfected cells and is blocked by the immunising peptide.
References for Anti-FOXP3 antibody (ab10563)
This product has been referenced in:
- Kosmidis Met al. Immunosuppression affects CD4+ mRNA expression and induces Th2 dominance in the microenvironment of cutaneous squamous cell carcinoma in organ transplant recipients. J Immunother 33:538-46 (2010). IHC-P; Human.Read more (PubMed: 20463594) »
- Chen YTet al. Interleukin-1 as a Phenotypic Immunomodulator in Keratinizing Squamous Metaplasia of the Ocular Surface in Sjogren's Syndrome. Am J Pathol 177:1333-43 (2010). IHC-P.Read more (PubMed: 20696775) »
See all 19 publications for this product
Publishing research using ab10563? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"