True-Nuclear™ One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP)

Pricing & Availability
Clone
MF-14 (See other available formats)
Other Names
Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
1_MF-14_True-Nuclear_AF488_1SKit_060915
C57BL/6 splenocytes were stained with True-Nuclear™ One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP).
  • 1_MF-14_True-Nuclear_AF488_1SKit_060915
    C57BL/6 splenocytes were stained with True-Nuclear™ One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP).
  • 2_MF-14_True-Nuclear_AF488_1SKit_060915
Cat # Size Price Quantity Avail. Save
136803 25 tests $500
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Description

FOXP3 is a 47 kD transcription factor, also known as Forkhead box protein P3, Scurfin, JM2, or IPEX. It is proposed to be a master regulatory gene and more specific marker of T regulatory cells than most cell surface markers (such as CD4 and CD25). Transduced expression of FOXP3 in CD4+/CD25- cells has been shown to induce GITR, CD103, and CTLA4 and impart a T regulatory cell phenotype. FOXP3 is mutated in X-linked autoimmunity-allergic dysregulation syndrome (XLAAD or IPEX) in humans and in "scurfy" mice. Overexpression of FOXP3 has been shown to lead to a hypoactive immune state suggesting that this transcriptional factor is a central regulator of T cell activity. In human, unlike in mouse, two isoforms of FOXP3 have been reported: one (FOXP3) corresponding to the canonical full-length sequence; the other (FOXP3 δ2) lacking exon 2. The 150D monoclonal antibody reacts with human, mouse and rat FOXP3. The 150D antibody recognizes FOXP3 epitope encoded by exon 2.

Product Details
Technical data sheet

Product Details

Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Concentration
Lot-specific (please contact technical support for concentration and total µg amount, or use our Lookup tool if you have a lot number.)
Storage & Handling
This kit is guaranteed for six months. Upon receipt, store between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC, ICFC - Quality tested

Recommended Usage

Materials Provided:
1. Alexa Fluor® 488 anti-mouse FOXP3/CD25 PE/CD4 PerCP antibody cocktail - 25 tests
2. Alexa Fluor® 488 Rat IgG2b, ? isotype control/CD25 PE/CD4 PerCP antibody cocktail - 25 tests
3. True-Nuclear™ Transcription Buffer Set - 120 tests (Cat. No. 424401)

Materials not included:
1. Cell Staining Buffer (Cat. No. 420201)
2. Single Color Compensation Controls

Immunofluorescence Staining Procedures:

1. Aliquot 100 µL of target cells to each tube.
2. Add 1 mL of the Transcription Factor 1X Fix solution to each tube, vortex, and incubate at room temperature in the dark for 45-60 minutes.
3. Without washing, add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
4. Centifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
5. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
6. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
7. Resuspend the cell pellet in 100 µL of the Transcription Factor 1X Perm Buffer.
8. Add 20 µL of Alexa Fluor® 488 anti-mouse FOXP3/CD25 PE/CD4 PerCP antibody cocktail or 20 µL of Alexa Fluor® 488 rat IgG2b, ? isotype control/CD25 PE/CD4 PerCP antibody cocktail into the appropriate tubes. Incubate in the dark at room temperature for at least 30 minutes.
9. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
10. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
11. Add 2 mL of the cell staining buffer.
12. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
13. Resuspend in 0.5 mL cell staining buffer and then acquire tubes on a flow cytometer.

Caution: The True-Nuclear™ Transcription Factor Buffer Set contains paraformaldehyde, which is toxic and mutagenic. Please handle with caution. Wear gloves, lab coats, and necessary protection to avoid direct contact.

NOTE: For flow cytometric staining with this clone, True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) offers improved staining and is highly recommended over the Foxp3/Perm Buffer Set (Cat. No. 421403).

Application Notes

NOTE: For flow cytometric staining with this clone, True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) offers improved staining and is highly recommended.

Application References

(PubMed link indicates BioLegend citation)
  1. Ono M, et al. 2007. Nature 446:685.
  2. Hori S, et al. 2003. Science 299:1057.
  3. Fontenot JD, et al. 2003 Nature Immunol 4:330.
  4. Fallarino F, et al. 2009. J. Immunol. 183:6033. PubMed
  5. Barber A, et al. 2009 J. Immunol. 183:6939. PubMed
  6. Nakashima H, et al. 2010. J. Immunol. 184:4637. PubMed
Product Citations
  1. Peng K, et al. 2017. PLoS One.. 10.1371/journal.pone.0183271. PubMed
Publication Library

Antigen Details

Structure
50-55 kd protein. Forkhead/winged-helix transcription factor family, contains zinc finger and forkhead domains.
Distribution

Nuclear; expressed in Treg cells.

Function
Master regulatory gene in Treg cell development, crucial for immune homeostasis.
Interaction
Interacts with DNA
Cell Type
Tregs
Biology Area
Immunology, Transcription Factors
Molecular Family
Nuclear Markers, CD Molecules
Antigen References

1. Ono M, et al. 2007. Nature 446:685.
2. Hori S, et al. 2003. Science 299:1057.
3. Fontenot JD, et al. 2003 Nature Immunol 4:330.
4. Fallarino F, et al. 2009. J. Immunol. 183:6033.
5. Barber A, et al. 2009 J. Immunol. 183:6939.
6. Nakashima H, et al. 2010. J. Immunol. 184:4637.

Regulation
Present at high level in T reg cells. Induced by T cell activation.
Gene ID
20371 View all products for this Gene ID
UniProt
View information about FOXP3 on UniProt.org

Related FAQs

Can FOXP3 be costained with cytokines?

The larger holes created by the nuclear permeabilization required for FOXP3 may allow cytokines to leak out of the cell, making it harder to detect lowly-expressed cytokines. You may have to use a control where the cells are only permeabilized through the cell membrane.

Can I stain whole blood with anti-FOXP3 using your Foxp3 staining kit?

It is not recommended. It is best to use PBMCs for this testing.

How stable is PerCP/Cy5.5 tandem as compared to PerCP alone?
PerCP/Cy5.5 is quite photostable and also better than PerCP alone in withstanding fixation.
Go To Top Version: 2    Revision Date: 05/26/2017

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