Biolegend, 421402, FOXP3 Perm Buffer (10x), 25ml

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.
25ml
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 Perform cell surface staining as described in Cell Surface Immunofluorescence Staining Protocol. 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). 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. (Optional stopping point) Resuspend cells in 1 mL of Cyto-Last™ Buffer (recommended) (Cat. No. 422501) or Cell Staining Buffer (Cat. No. 420201) and store at 4°C. When ready to continue, centrifuge at 250 × g for 5 minutes. Discard the supernatant and proceed to the next step. 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. Wash once with 1 ml of 1X FOXP3 Perm Buffer. 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. Add an appropriate amount of fluorochrome-conjugated anti-FOXP3 antibody and incubate at room temperature in the dark for 30 minutes. Wash twice with Cell Staining Buffer, and resuspend in 0.5 ml of Cell Staining Buffer, then analyze with a flow cytometer.
Product Citations: Cheng Y, et al. 2016. J Immunol. 196: 924 - 932. PubMed Sapski S, et al. 2020. Cancer Immunol Immunother. 2291:69. PubMed Jia C, et al. 2015. Fish Shellfish Immunol. 45: 300-306. PubMed Li H, et al. 2021. Adv Sci (Weinh). 2001596:8. PubMed Wei C, et al. 2016. Cell Death Dis. 7:e2489. PubMed
Biology Area: Transcription Factors
Molecular Family: Nuclear Markers
Gene ID: NA
Regulatory Status: RUO
Other Names: FOXP3 Permeabilization Buffer
Q: Can I stain whole blood with anti-FOXP3 using your Foxp3 staining kit?
A: It is not recommended. It is best to use PBMCs for this testing.
Q: Can FOXP3 be costained with cytokines?
A: 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.