True-Nuclear™ Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD4 APC/CD25 PE)

Pricing & Availability
Clone
150D (See other available formats)
Other Names
Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
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Product Citations
publications
TrueNuclear_MU_Treg_FlowKit_FOXP3A488_CD4APC_CD25PE_1_052616
BALB/c mouse splenocytes were stained with True-Nuclear™ Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD4 APC/CD25 PE)
  • TrueNuclear_MU_Treg_FlowKit_FOXP3A488_CD4APC_CD25PE_1_052616
    BALB/c mouse splenocytes were stained with True-Nuclear™ Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD4 APC/CD25 PE)
  • TrueNuclear_MU_Treg_FlowKit_FOXP3A488_CD4APC_CD25PE_2_052616
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Cat # Size Price Quantity Avail. Save
320029 25 tests $450
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Description

T regulatory (Treg) cells are a subset of T lymphocytes which is characterized by CD4+/CD25+/FOXP3+. These naturally occurring Treg cells originate in the thymus, and comprise 2-10% of peripheral CD4+ T cells. It has been shown that Treg cells are able to inhibit T cells proliferation and cytokine production and play critical roles in preventing autoimmunity as well as in controlling tumor immunity and transplantation tolerance. Impaired Treg function or Treg cell deficiency will develop variety of autoimmune diseases, while higher frequency of Treg cells will cause hypo-immune response to pathogens.


BioLegend's True-Nuclear™ Mouse Treg Flow™ Kit is designed and formulated specifically for immunofluroscence staining and flow cytometric analysis of mouse Treg cells in a mixed lymphocyte population. This kit is composed of fluorochrome conjugated anti-mouse CD4, CD25, FOXP3 antibodies, and the critical buffers. It is easy to use for identification of Treg cells.

Product Details
Technical data sheet

Product Details

Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Mouse
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

Application Notes

Materials Provided:
1.
Alexa Fluor® 488 anti-mouse FOXP3 - 25 tests
2. Alexa Fluor® 488 Mouse IgG1, κ isotype control - 25 tests
3. True-Nuclear™ Transcription Factor Buffer Set - 120 tests (Cat. No. 424401)
4. anti-mouse CD4 APC/CD25 PE Cocktail - 50 tests

Materials Not Included:
1.
Cell Staining Buffer (Cat. No. 420201)
2. Single color compensation controls

Immunofluorescence Staining Procedures:

1. Perform cell surface staining as described in BioLegend's Cell Surface Immunofluorescence Staining Protocol. Add 20 µL of the anti-mouse CD4 APC/CD25 PE cocktail to each tube and incubate in the dark for 20 minutes.
2. Add 2 mL of the cell staining buffer, centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
3. Repeat Step 2, for a total of two washes.
4. 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.
5. Without washing, 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. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
8. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
9. Resuspend the cell pellet in 100 µl of the Transcription Factor 1XPerm Buffer.
10. Add 5 µL of Alexa Fluor® 488 anti-mouse FOXP3 antibody or 5 µL of Alexa Fluor® 488 mouse IgG1, κ isotype control into the appropriate tubes. Incubate in the dark at room temperature for at least 30 minutes.
11. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
12. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
13. Add 2 mL of the cell staining buffer.
14. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
15. 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. Roncador G, et al. 2005 Eur. J. Immunol. 35:1681.
  2. Mayack. S,et al. 2006. J. Immunol.176:2059. J. Immunol
  3. Yang ZZ, et al. 2006. Blood 107:3639.
  4. Gavin MA, et al. 2006. P. Natl. Acad. Sci. USA 103:6659.
  5. Groh V, et al. 2006. Nature Immunology 7:755.
  6. Lewkowicz P, et al. 2006 J. Immunol. 177:7155.
  7. Luke PPW, et al. 2006. Amer. J. Transplant. 6(9):2023.
  8. Bamias G, et al. 2007. J. Immunol. 178:1809.
  9. Valencia X, et al. 2007. J. Immunol. 178:2579.PubMed
  10. Davidson TS, et al. 2007. J. Immunol. 178:4022.
  11. MacDonald K PA, et al. 2007. Blood doi:10.1182/blood-2007-01-067249.
  12. Jaffar Z, et al. 2007. J. Immunol. 179:6193.
  13. Müller M, et al. 2007. J. Immunol. 179:2774.
  14. Jordan JM,et al. 2008.Infect Human. 76:3717. PubMed
  15. Golovina TN,et al. 2008. J. Immunol. 181:2855. PubMed
  16. Fallarino F, et al. 2009. J. Exp Med. 206:2511. PubMed
  17. Banham Alison, et al. 2009. Vet Immunol and Immunop 127.3-4:376-381
  18. Klunker S, et al. 2009. J. Exp Med. PubMed
  19. Haque A, et al. 2010. J. Immunol. 184:2583. PubMed
  20. Liu Y, et al. 2012. Food Chem Toxicol. 50:1920. PubMed

Antigen Details

Distribution

T regulatory cells

Cell Type
Tregs
Biology Area
Cell Biology, Immunology, Transcription Factors
Molecular Family
CD Molecules, Nuclear Markers
Antigen References

1. Hori S, et al. 2003. Science 299:1057.
2. Fontenot JD, et al. 2003. Nature Immunol. 4:330.
3. Ferguson PJ, et al. 2000. Am. J. Med. Genet. 90:390.
4. Bennett CL, et al. 2001. Nature Genet. 27:20.
5. Allan SE, et al. 2005. J. Clin. Invest. 115:3276.

Gene ID
20371 View all products for this Gene ID 317382 View all products for this Gene ID 50943 View all products for this Gene ID
UniProt
View information about Treg Flow Kit 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.

What type of PE do you use in your conjugates?
We use R-PE in our conjugates.
Go To Top Version: 3    Revision Date: 12/15/2016

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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