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FOXP3 Perm Buffer (10x)

Pricing & Availability
Regulatory Status
RUO
Other Names
FOXP3 Permeabilization Buffer
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Product Citations
publications
A_FoxP3_True-Nuclear_Flow_Chart_060117
Whole blood was stained with human CD3 APC/Cyanine7 (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/Cyanine7 channels increased significantly for the FoxP3 Fix/Perm buffer set versus the True-Nuclear™ transcription factor buffer set for both APC/Cyanine7 and APC/Fire™ 750.
  • A_FoxP3_True-Nuclear_Flow_Chart_060117
    Whole blood was stained with human CD3 APC/Cyanine7 (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/Cyanine7 channels increased significantly for the FoxP3 Fix/Perm buffer set versus the True-Nuclear™ transcription factor buffer set for both APC/Cyanine7 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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421402 25 mL £49
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Description

This 25 mL FOXP3 Perm Buffer is 10X concentrated and designed for use with the FOXP3 Fix/Perm Buffer (Cat. No. 421401). It should 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 Perm buffer (10X) to nine (9) parts PBS.

Warning: The FOXP3 Buffer Set can have a deleterious effect on tandem fluorophores (particularly APC/Cyanine7) 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
Store between 2°C and 8°C. Do not freeze.

To obtain lot-specific expiration date, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.)
Application

ICFC - Quality tested
Recommended Usage

The FOXP3 Perm Buffer (10x) must be diluted to a 1X working solution with PBS prior to staining procedures. Dilute 1 part FOXP3 Perm Buffer (10x) to 9 parts PBS (10 mM sodium phosphate, 150 mM sodium chloride, pH 7.2).

NOTE: FOXP3 Perm Buffer (10x) may exhibit crystallization or precipitation when stored at 2-8°.  This is normal and does not affect the buffer's performance. Heavy precipitation observed after dilution to a 1x solution can be cleared by filtration.

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 Procedure

  1. Perform cell surface staining as described in Cell Surface Immunofluorescence Staining Protocol.
  2. Prepare a 1x working solution of FOXP3 Fix/Perm Buffer by diluting 1 part FOXP3 Fix/Perm Buffer (4x) (Cat. No. 421401) in 3 parts PBS (10 mM sodium phosphate, 150 mM sodium chloride, pH 7.2).  
  3. Add 1 ml of 1X FOXP3 Fix/Perm solution to each sample. Vortex and incubate at room temperature in the dark for 20 minutes. Spin at 250 x g for 5 minutes and discard the supernatant.
  4. Add 1 ml of Cell Staining Buffer (Cat. No. 420201) to wash cells. Spin at 250 x g for 5 minutes and discard the supernatant.
  5. Prepare a 1x working solution of FOXP3 Perm Buffer by diluting 1 part FOXP3 Perm Buffer (10x) (Cat. No. 421402) to 9 parts PBS (10 mM sodium phosphate, 150 mM sodium chloride, pH 7.2).

NOTE: FOXP3 Perm Buffer (10x) may exhibit crystallization or precipitation when stored at 2-8°.  This is normal and does not affect the buffer's performance. Heavy precipitation observed after dilution to a 1x solution can be cleared by filtration.

  1. Wash once with 1 ml of 1X FOXP3 Perm Buffer.
  2. Resuspend cells in 1 ml of 1X 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 μL of 1X FOXP3 Perm Buffer.
  3. Add an appropriate amount of fluorochrome-conjugated anti-FOXP3 antibody and incubate at room temperature in the dark for 30 minutes.
  4. Wash twice with Cell Staining Buffer, and resuspend in 0.5 ml of Cell Staining Buffer, then analyze with a flow cytometer.
Product Citations
  1. Cheng Y, et al. 2016. J Immunol. 196: 924 - 932. PubMed
  2. Sapski S, et al. 2020. Cancer Immunol Immunother. 2291:69. PubMed
  3. Jia C, et al. 2015. Fish Shellfish Immunol. 45: 300-306. PubMed
  4. Li H, et al. 2021. Adv Sci (Weinh). 2001596:8. PubMed
  5. Wei C, et al. 2016. Cell Death Dis. 7:e2489. PubMed

Antigen Details

Biology Area
Transcription Factors
Molecular Family
Nuclear Markers
Gene ID
NA

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.

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.

Go To Top Version: 5    Revision Date: 09/16/2024

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.

 

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