- Other Names
- FOXP3 Permeabilization Buffer
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This 25 ml FOXP3 Perm Buffer is 10X concentrated and designed for use with the FOXP3 Fix/Perm Buffer (cat# 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/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.
- Storage & Handling
Store 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.
ICFC - Quality tested
- Recommended Usage
The FOXP3 Perm Buffer (10x) 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.
NOTE: The FOXP3 Perm Buffer (10x) may have crystalization or precipitation oberserved when it is stored at 2-8°C; however, it's normal and does not affect the buffer performance. If there is a heavy precipitation observed after diluted to 1X working solution, it can be filtered to clarify the solution.
- 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.
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- Biology Area
- Transcription Factors
- Molecular Family
- Nuclear Markers
- Gene ID
- 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.