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One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP)
One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP)
136801 25 tests $500.00     
Product Details
Clone: MF-14
Reactivity: Mouse
Storage & Handling: This kit is guaranteed for three months.
Application:

FC, ICFC - Quality tested
COA:
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Application
Notes:

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. FOXP3 Fix/Perm Buffer Set, 100 tests Cat. No. 421403

Materials not included:
Cell Staining Buffer (Cat. No. 420201)

Immunofluorescence Staining Procedures:

Centrifugation steps: perform at 250Xg for 5min
Incubation steps: perform at room temperature.

1. Prepare 1X working solutions of FOXP3 Fix/Perm buffer and FOXP3 Perm buffer prior to staining. The FOXP3 Fix/Perm buffer (4X) must be freshly diluted by diluting one (1) part FOPX3 Fix/Perm buffer (4X) with three (3) parts PBS. The FOXP3 Perm buffer (10X) should be diluted by diluting one (1) part FOXP3 Perm buffer (10X) with nine (9) parts PBS.

NOTE: The FOXP3 Perm buffer (10X) may have crystalization or precipitation observed when it is stored at 2-8°C, however, it is normal and does not affect the buffer performance. If there is heavy precipitation observed after diluted to 1X working solution, it can be filtered to clarify the solution.
Caution: The FOXP3 Fix/Perm buffer contains paraformaldehyde, which is toxic and mutagenic. Please wear necessary protection to avoid direct body contact.

2. Suspend the cells of interest at 1X107/ml  in BioLegend's cell staining buffer (Cat. No. 420201).
3. Distribute 100 μl of the cell suspension into each FACS tube.
4. Add 1 ml of 1X BioLegend's FOXP3 Fix/Perm solution to each tube, vortex and incubate for 20 minutes; centrifuge, then discard the supernatant.
5. Wash: resuspend cells in 2ml/tube cell staining buffer; centrifuge, then discard the supernatant.
6. Wash: resuspend in 1ml 1X BioLegend's FOXP3 Perm buffer; centrifuge, then discard the supernatent.
7. Resuspend cells in 1ml 1X BioLegend's FOXP3 Perm buffer, incubate in the dark for 15 minutes; centrifuge, then discard the supernatant.
8. Add 20 µl Alexa Fluor® 488 anti-mouse FOXP3/CD25 PE/CD4 PerCP antibody cocktail or 20 µl Alexa Fluor® 488 rat IgG2b, κ isotype control/CD25 PE/CD4 PerCP antibody cocktail into appropriate tube, and resuspend the cell pellet by gentle vortex. Incubate in the dark for 30 minutes.
9. Wash twice with cell staining buffer (see step 5) and resuspend in 0.5ml cell staining buffer then analyze with flow cytometer using appropriate instrument settings.
Application
References:

1. Ono M, et al.:Nature 2007 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
Splenocytes from C57BL/6 mouse stained
Splenocytes from C57BL/6 mouse stained with One Step Staining Mouse Treg FlowTM Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP )


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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 One Step Staining Mouse Treg Flow™ Kit is designed and formulated specifically for immunofluorescence 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 and FOXP3 antibodies and the critical buffers. It is an one-step staining kit for labeling both surface and intracellular antigens of Treg cells.  
Other Names: Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2, T regulatory cells, Regulatory T cells
Structure: Treg Flow Kit
Distribution: T regulatory cells
Antigen
References:

1. OnoM, et al. Nature 2007 446:685
GeneID: 20371
Latest Publications: View the latest FOXP3 articles on HighwirePress.com
UniProt: View information about FOXP3 on UniProt.org
Keywords: One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP), MF-14, Kit, Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2, T regulatory cells, Regulatory T cells, Mouse, Immunology, Antibodies
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Compare Data Across All Formats
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.
  • Purified anti-mouse FOXP3
    Western blot analysis of extracts

    Western blot analysis of extracts from untransfected 293T cells (Lane 1) or 293T cells transfected with a plasmid encoding mouse FOXP3 protein (Lane 2), using anti-FOXP3, clone MF-14.

  • PE anti-mouse FOXP3
    C57BL/6 splenocytes surface stained with

    C57BL/6 splenocytes surface stained with CD4 (GK1.5) APC and then intracellularly stained MF-14 PE

  • Alexa Fluor® 488 anti-mouse FOXP3
    C57BL/6 splenocytes were surface stained

    C57BL/6 splenocytes were surface stained with CD4 (GK1.5) APC and MF-14 Alexa Fluor® 488

  • Alexa Fluor® 647 anti-mouse FOXP3
    C57BL/6 splenocytes surface stained with

    C57BL/6 splenocytes surface stained with CD4 (GK1.5) PE and then intracellularly stained with MF-14 Alexa Fluor® 647

  • Pacific Blue™ anti-mouse FOXP3
    C57BL/6 splenocytes surface stained with

    C57BL/6 splenocytes surface stained with CD4 (GK1.5) PE and then intracellularly stained with MF-14 Pacific Blue™

  • One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP)
    Splenocytes from C57BL/6 mouse stained

    Splenocytes from C57BL/6 mouse stained with One Step Staining Mouse Treg FlowTM Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP )

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