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True-Nuclear™ One Step Mouse Treg Flow Kit

Pricing & Availability
Clone
MF-14 (See other available formats)
Regulatory Status
RUO
Other Names
Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
Isotype
Rat IgG2b, κ
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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
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136803 25 tests $586
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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.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
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

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, κ 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 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
  2. Lu Y, et al. 2018. Cancer Cell. 33:1048. PubMed
RRID
AB_3097010 (BioLegend Cat. No. 136803)

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
CD Molecules, Nuclear Markers
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.

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 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.

Go To Top Version: 2    Revision Date: 05/26/2017

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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