FOXP3 Fix/Perm Buffer (4x)

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Other Names
FOXP3 Fixation/Permeabilization Buffer
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Product Citations
publications
A_FoxP3_True-Nuclear_Flow_Chart_060117
Whole blood was stained with human CD3 APC/Cy7 (blue and red) or APC/Fire™ 750 (green and purple), then lysed, washed and fixed under the following conditions: A) Fixed with FOXP3 fix buffer followed by permeabilization with FOXP3 perm buffer (purple and red). B) Fixed with True-Nuclear™ fix buffer, then permeabilized with True-Nuclear™ perm buffer (green and blue). Compensation requirements between the APC and APC/Cy7 channels increased significantly for the FoxP3 Fix/Perm buffer set versus the True-Nuclear™ transcription factor buffer set for both APC/Cy7 and APC/Fire™ 750.
  • A_FoxP3_True-Nuclear_Flow_Chart_060117
    Whole blood was stained with human CD3 APC/Cy7 (blue and red) or APC/Fire™ 750 (green and purple), then lysed, washed and fixed under the following conditions: A) Fixed with FOXP3 fix buffer followed by permeabilization with FOXP3 perm buffer (purple and red). B) Fixed with True-Nuclear™ fix buffer, then permeabilized with True-Nuclear™ perm buffer (green and blue). Compensation requirements between the APC and APC/Cy7 channels increased significantly for the FoxP3 Fix/Perm buffer set versus the True-Nuclear™ transcription factor buffer set for both APC/Cy7 and APC/Fire™ 750.
  • B_FoxP3_True-Nuclear_BALBC_Plots_053117
    BALB/c spleen cells were cultured for four days in presence of LPS. Then the cells were stained with CD45R/B220 PE, followed by fixation and permeabilization with either FoxP3 Fix/Perm Buffer (top) or True-Nuclear™ Transcription Factor Buffer Set (bottom), and staining with Blimp-1 (clone 5E7) Alexa Fluor® 647. Data shown was gated on live cells using Zombie Aqua™ fixable viability dye.
  • C_FoxP3_True-Nuclear_HuPMBC_Plots_053117
    Human peripheral blood lymphocytes were surface stained with CD4 APC and then treated with True-Nuclear™ Transcription Factor Buffer Set (Cat# 424401). Cells were then stained with FOXP3 (clone 206D) Brilliant Violet 421™ (top) or mouse IgG1, κ Brilliant Violet 421™ isotype control (bottom).
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421401 30 ml £40
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Description

BioLegend's FOXP3 Fix/Perm Buffer (4x) has been specially formulated for intracellular staining of FOXP3 with minimum effect on the surface fluorochrome staining. The FOXP3 Fix/Perm buffer(4X) must be freshly diluted to 1X working solution with PBS (10 mM sodium phosphate, 150 mM sodium chloride, pH 7.2) prior to staining procedures. e.g. Dilute one (1) part FOXP3 Fix/Perm buffer(4X) to three (3) parts PBS.

Warning: The FOXP3 Buffer Set can have a deleterious effect on tandem fluorophores (particularly APC/Cy7) in a multicolor assay, causing an increase in compensation into the APC channel.  If your panel contains a surface stain with an antibody conjugated to a tandem fluorophore, consider using the True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) instead, which provides a clearer resolution of FoxP3 as well as other nuclear transcription factors. Please refer to the data provided here.

Caution: The FOXP3 Fix/Perm buffer contains paraformaldehyde, which is toxigenic and mutagenic. Please handle with caution and wear gloves, lab coat and necessary protection to avoid direct body contact.

Product Details
Technical Data Sheet (pdf)

Product Details

Storage & Handling
This solution is guaranteed for three months. It should be stored between 2°C and 8°C. Do not freeze.

This product has a shelf-life of 12 months or less. Please contact our technical support team for lot specific CoA and expiration date inquiries of this product.
Application

ICFC - Quality tested

Recommended Usage

The FOXP3 Fix/Perm buffer(4X) must be diluted to (1X) working solution with PBS (10 mM sodium phosphate, 150 mM sodium chloride, pH 7.2) prior to staining procedures. e.g. Dilute one (1) part FOXP3 Fix/Perm buffer(4X) to three (3) parts PBS.

Application Notes

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

FOXP3 Intracellular Staining Procedures:
1. Perform cell surface staining as described in BioLegend's Cell Surface Immunofluorescence Staining Protocol.
2. Add 1 ml of 1X BioLegend's FOXP3 Fix/Perm solution to each tube, vortex and incubate at room temperature in the dark for 20 minutes, then spin down the cells and remove the supernatant.
3. Wash once with cell staining buffer (Cat. No. 420201). Spin at 250Xg for 5 minutes and remove the supernatant.
4. Wash once with 1ml 1X BioLegend's FOXP3 Perm buffer.
5. Re-suspend cells in 1ml 1X BioLegend's FOXP3 Perm buffer, incubate at room temperature in the dark for 15 minutes, spin down cells and discard the supernatant, then resuspend the pellet in 100 ul of 1X BioLegend's FOXP3 Perm buffer.
6. Add appropriate amount of flurochrome conjugated anti-FOXP3 antibody and incubate at room temperature in the dark for 30 minutes.
7. Wash twice with cell staining buffer, and resuspend in 0.5ml cell staining buffer then analyze with flow cytometer with appropriate instrument setting.

Product Citations
  1. He T, et al. 2015. J Virol. 89: 9616-9630. PubMed
  2. Kaufmann U, et al. 2016. J Immunol. 196: 573 - 585. PubMed
  3. Albeituni S, et al. 2016. J Immunol. 196: 2167 - 2180. PubMed
  4. Wei C, et al. 2016. Cell Death Dis. 7:e2489. PubMed
Publication Library

Antigen Details

Biology Area
Transcription Factors
Molecular Family
Nuclear Markers
Gene ID
NA

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.

Go To Top Version: 2    Revision Date: 02/03/2015

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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