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FOXP3 Fix/Perm Buffer Set FOXP3 Fix/Perm Buffer Set

Pricing & Availability
Regulatory Status
RUO
Other Names
FOXP3 Fixation/Permeabilization Buffer
Ave. Rating
Submit a Review
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. No. 424401). Cells were then stained with FOXP3 (clone 206D) Brilliant Violet 421™ (top) or mouse IgG1, κ Brilliant Violet 421™ isotype control (bottom).
Cat # Size Price Quantity Check Availability Save
421403 100 tests NOK1164
Check Availability


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Request Bulk Quote
Description

BioLegend's FOXP3 Fix/Perm Buffer Set has been specially formulated for intracellular staining FOXP3 with minimum effect on the surface fluorochrome staining. It contains one 30 ml 4X concentrated FOXP3 Fix/Perm buffer (Cat. No. 421401) and one 25 mL 10X concentrated FOXP3 Perm buffer (Cat. No. 421402). 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. 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.

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.)

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 a 1X working solution with PBS  prior to staining procedures. Dilute 1 part FOXP3 Fix/Perm Buffer (4x) to 3 parts PBS (10 mM sodium phosphate, 150 mM sodium chloride, pH 7.2). 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.

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.
Application References
  1. Yang ZZ, et al. 2006. Blood 107:3639. PubMed
  2. Groh V, et al. 2006. Nat. Immunol. 7:755. PubMed
  3. Bamias G, et al. 2007. J. Immunol. 178:1809. PubMed
  4. MacDonald K PA, et al. 2007. Blood doi:10.1182/blood-2007-01-067249. PubMed
  5. Müller M, et al. 2007. J. Immunol. 179:2774.
  6. Dang Y,et al.2007.Clin Cancer Res.13:1883. PubMed
  7. Basu S, et al. 2008. J. Immunol. 180:5974. PubMed
  8. Takamura S, et al. 2010. J. Immunol. 184:4696. PubMed
  9. Beavis, PA., et al. 2011. PNAS. 108:16717. PubMed
  10. Kling J, et al. 2013. Exp Parasitol. 129:270. PubMed
  11. Troutman TD, et al. 2012. PNAS. 109:273. PubMed
  12. Faustman DL, et al. 2012. PLoS One. 7:e41756. PubMed
  13. Liu Y, et al. 2012. Food Chem Toxicol. 50:1920. PubMed
  14. Kawamoto N, et al. 2013. Int Immunol. 25:53. PubMed
  15. Morandi B, et al. 2013. J. Immunol. 191:4858. PubMed
  16. Shiba T, et al. 2014. Toxicol Appl Pharmacol. 274:191. PubMed
  17. Cook KW, et al. 2014. Gut. PubMed
  18. Shibuya KC, et al. 2014. PLoS One. 9:96565. PubMed
  19. Cecil DL, et al. 2014. Cancer Res. 74:2710. PubMed
  20. Linnerbauer S, et al. 2014. PLoS Pathog. 10:1004068. PubMed
  21. Basu S, et al. 2015. J Leukoc Biol. 97:279. PubMed
Product Citations
  1. Shibuya K, et al. 2014. PLoS One. 9:96565. PubMed
  2. Williams JW, et al. 2020. Nat Immunol. 1194:21. PubMed
  3. Danileviciute E, et al. 2022. Nat Metab. 4:589. PubMed
  4. Freimer JW, et al. 2022. Nat Genet. 54:1133. PubMed
  5. Li L, et al. 2022. Cancer Immunol Res. 10:1475. PubMed
  6. Fetit R, et al. 2023. Development. 150:. PubMed
  7. Jin J, et al. 2023. Nat Aging. 3:600. PubMed
  8. Chiu H, et al. 2021. iScience. 24:102748. PubMed
  9. Vella JL, et al. 2021. Life Sci Alliance. 4:. PubMed
  10. Jin J, et al. 2021. Sci Immunol. 6:. PubMed
  11. Hippen KL, et al. 2021. Cytotherapy. 23:704. PubMed
  12. Ren J, et al. 2022. Proc Natl Acad Sci U S A. 119:e2117401119. PubMed
  13. Guttman O, et al. 2022. J Cell Biol. 221: . PubMed
  14. Wang Z, et al. 2022. Proc Natl Acad Sci U S A. 119:e2201899119. PubMed
  15. Zhou Y, et al. 2022. Evid Based Complement Alternat Med. 2022:2197763. PubMed
  16. Hajjar S, et al. 2022. Cell Death Differ. 29:585. PubMed
  17. Yang D, et al. 2016. PLoS Negl Trop Dis. 10: 0004335. PubMed
  18. Singh AK, et al. 2017. JCI Insight. 2:e94275. PubMed
  19. Zbinden A, et al. 2021. Adv Sci (Weinh). 2002500:8. PubMed
  20. Mizraji G, et al. 2017. Cell Rep. 18(2):419-431. PubMed
  21. Puc I, et al. 2022. Int J Mol Sci. 23:. PubMed
  22. Song J, et al. 2022. Clin Transl Med. 12:e992. PubMed
  23. Zemek RM, et al. 2020. Nat Protoc. 15:1628. PubMed
  24. Troutman T, et al. 2012. Proc Natl Acad Sci U S A. 109:273. PubMed
  25. Kawamoto N, et al. 2013. Int Immunol. 25:53. PubMed
  26. Kamigaki T, et al. 2015. Anticancer Res. 35: 4535-4543. PubMed
  27. Fernandez SV, et al. 2020. Breast Cancer Res. 1.009722222. PubMed
  28. Cook K, et al. 2014. Gut. . PubMed
  29. Bamias G, et al. 2007. J Immunol. 178:1809. PubMed
  30. Dang Y, et al. 2007. Clin Cancer Res . 1.932638889. PubMed
  31. Yang Z, et al. 2006. Blood. 107:3639. PubMed
  32. Morandi B, et al. 2013. J Immunol. 191:4858. PubMed
  33. Satoh M, et al. 2016. Sci Rep. 6:28473. PubMed
  34. Zhao XY, et al. 2020. Sci Adv. 6:eaay6191. PubMed
  35. Bankoti R, et al. 2017. Sci Rep. 10.1038/s41598-017-12171-3. PubMed
  36. Basu S, et al. 2015. J Leukoc Biol. 97:279. PubMed
  37. Haller M, et al. 2016. Diabetes. 65(12):3765-3775. PubMed
  38. Van Gool F, et al. 2019. Immunity. 50:362. PubMed
  39. Takamura S, et al. 2010. J Immunol. 184:4696. PubMed
  40. Brennan CA, et al. 2021. Gut Microbes. 13:1987780. PubMed
  41. Scur M, et al. 2022. Nat Commun. 13:7272. PubMed
  42. Greaney SK, et al. 2020. Cancer Immunol Res. 0.504166667. PubMed
  43. Matisz C, et al. 2017. Sci Rep. 7:40631. PubMed
  44. Pfirrmann V, et al. 2015. Cytotherapy. 17: 1139-1151. PubMed
  45. Cecil D, et al. 2014. Cancer Res. 74:2710. PubMed
  46. Liu Y, et al. 2012. Food Chem Toxicol. 50:1920. PubMed
  47. Beavis P, et al. 2011. Proc Natl Acad Sci U S A. 108:16717. PubMed
  48. Zhou Q, et al. 2015. J Immunol. 195: 2493-250. PubMed
  49. Huang S, et al. 2016. Immunol Cell Biol. 10.1038/icb.2016.75. PubMed
  50. Basu S, et al. 2008. J Immunol. 180:5794. PubMed
  51. Luo G, et al. 2021. Sci Rep. 11:7841. PubMed
  52. Keefe RC, et al. 2021. Sci Rep. 11:14933. PubMed
  53. Kwek S, et al. 2015. Cancer Immunol Res. 3: 1008-1016. PubMed
  54. Shiba T, et al. 2014. Toxicol Appl Pharmacol. 274:191. PubMed
  55. de Winde CM, et al. 2021. J Cell Sci. 134: . PubMed
  56. Li H, et al. 2015. Reproduction. 150: 417-427. PubMed
  57. Andersen K, et al. 2017. J Am Soc Nephrol. 28:76. PubMed
  58. Oliveira G, et al. 2015. Sci Transl Med. 7: 317ra198. PubMed
  59. Urueña C, et al. 2022. Front Med (Lausanne). 9:991873. PubMed
  60. Reuschl AK, et al. 2022. Cell Rep. 39:110650. PubMed
  61. Chiaro J, et al. 2021. Cancer Immunol Res. 9:981. PubMed
  62. Solleiro-Villavicencio H, et al. 2015. Clin Immunol. . PubMed
  63. Martini E, et al. 2016. Cell Rep. 14:1062-1073. PubMed
  64. Jacobs H, et al. 2016. J Immunol. 196: 3525 - 3531. PubMed
  65. MacDonald K, et al. 2007. Blood . 109:5049. PubMed
  66. Méndez-Lagares G, et al. 2021. J Clin Invest. 131:e148542. PubMed
  67. Mishra P, et al. 2016. PLoS Negl Trop Dis. 10: 0004787. PubMed
  68. Weber A, et al. 2016. Infect Immun . 84: 1413 - 1423. PubMed
  69. Linnerbauer S, et al. 2014. PLoS Pathog. 10:1004068. PubMed
  70. Zheng D, et al. 2022. Front Immunol. 13:808347. PubMed
  71. Kling J, et al. 2011. Exp Parasitol. 129:270. PubMed
  72. Okagawa T, et al. 2016. Infect Immun . 84: 77 - 89. PubMed
  73. Namas R, et al. 2016. Lupus Sci Med. 3: e000148. PubMed
  74. Javed A, et al. 2015. PLoS One. 10: 0142086. PubMed
  75. Li Y, et al. 2015. J Immunol. 195: 1591 - 1598. PubMed
  76. Faustman D, et al. 2012. PLoS One. 7:e41756. PubMed
  77. Alizadeh A, et al. 2018. J Neuroinflammation. 15:53. PubMed
  78. Okano T, et al. 2017. Sci Rep. 7:42412. PubMed
  79. Aggarwal N, et al. 2021. Cell Rep. 37:110170. PubMed
  80. Shibuya K, et al. 2014. PLoS One. 9:96565. PubMed
  81. Williams JW, et al. 2020. Nat Immunol. 1194:21. PubMed
  82. Danileviciute E, et al. 2022. Nat Metab. 4:589. PubMed
  83. Freimer JW, et al. 2022. Nat Genet. 54:1133. PubMed
  84. Li L, et al. 2022. Cancer Immunol Res. 10:1475. PubMed
  85. Fetit R, et al. 2023. Development. 150:. PubMed
  86. Jin J, et al. 2023. Nat Aging. 3:600. PubMed
  87. Chiu H, et al. 2021. iScience. 24:102748. PubMed
  88. Vella JL, et al. 2021. Life Sci Alliance. 4:. PubMed
  89. Jin J, et al. 2021. Sci Immunol. 6:. PubMed
  90. Hippen KL, et al. 2021. Cytotherapy. 23:704. PubMed
  91. Ren J, et al. 2022. Proc Natl Acad Sci U S A. 119:e2117401119. PubMed
  92. Guttman O, et al. 2022. J Cell Biol. 221: . PubMed
  93. Wang Z, et al. 2022. Proc Natl Acad Sci U S A. 119:e2201899119. PubMed
  94. Zhou Y, et al. 2022. Evid Based Complement Alternat Med. 2022:2197763. PubMed
  95. Hajjar S, et al. 2022. Cell Death Differ. 29:585. PubMed
  96. Yang D, et al. 2016. PLoS Negl Trop Dis. 10: 0004335. PubMed
  97. Singh AK, et al. 2017. JCI Insight. 2:e94275. PubMed
  98. Zbinden A, et al. 2021. Adv Sci (Weinh). 2002500:8. PubMed
  99. Mizraji G, et al. 2017. Cell Rep. 18(2):419-431. PubMed
  100. Puc I, et al. 2022. Int J Mol Sci. 23:. PubMed
  101. Song J, et al. 2022. Clin Transl Med. 12:e992. PubMed
  102. Zemek RM, et al. 2020. Nat Protoc. 15:1628. PubMed
  103. Troutman T, et al. 2012. Proc Natl Acad Sci U S A. 109:273. PubMed
  104. Kawamoto N, et al. 2013. Int Immunol. 25:53. PubMed
  105. Kamigaki T, et al. 2015. Anticancer Res. 35: 4535-4543. PubMed
  106. Fernandez SV, et al. 2020. Breast Cancer Res. 1.009722222. PubMed
  107. Cook K, et al. 2014. Gut. . PubMed
  108. Bamias G, et al. 2007. J Immunol. 178:1809. PubMed
  109. Dang Y, et al. 2007. Clin Cancer Res . 1.932638889. PubMed
  110. Yang Z, et al. 2006. Blood. 107:3639. PubMed
  111. Morandi B, et al. 2013. J Immunol. 191:4858. PubMed
  112. Satoh M, et al. 2016. Sci Rep. 6:28473. PubMed
  113. Zhao XY, et al. 2020. Sci Adv. 6:eaay6191. PubMed
  114. Bankoti R, et al. 2017. Sci Rep. 10.1038/s41598-017-12171-3. PubMed
  115. Basu S, et al. 2015. J Leukoc Biol. 97:279. PubMed
  116. Haller M, et al. 2016. Diabetes. 65(12):3765-3775. PubMed
  117. Van Gool F, et al. 2019. Immunity. 50:362. PubMed
  118. Takamura S, et al. 2010. J Immunol. 184:4696. PubMed
  119. Brennan CA, et al. 2021. Gut Microbes. 13:1987780. PubMed
  120. Scur M, et al. 2022. Nat Commun. 13:7272. PubMed
  121. Greaney SK, et al. 2020. Cancer Immunol Res. 0.504166667. PubMed
  122. Matisz C, et al. 2017. Sci Rep. 7:40631. PubMed
  123. Pfirrmann V, et al. 2015. Cytotherapy. 17: 1139-1151. PubMed
  124. Cecil D, et al. 2014. Cancer Res. 74:2710. PubMed
  125. Liu Y, et al. 2012. Food Chem Toxicol. 50:1920. PubMed
  126. Beavis P, et al. 2011. Proc Natl Acad Sci U S A. 108:16717. PubMed
  127. Zhou Q, et al. 2015. J Immunol. 195: 2493-250. PubMed
  128. Huang S, et al. 2016. Immunol Cell Biol. 10.1038/icb.2016.75. PubMed
  129. Basu S, et al. 2008. J Immunol. 180:5794. PubMed
  130. Luo G, et al. 2021. Sci Rep. 11:7841. PubMed
  131. Keefe RC, et al. 2021. Sci Rep. 11:14933. PubMed
  132. Kwek S, et al. 2015. Cancer Immunol Res. 3: 1008-1016. PubMed
  133. Shiba T, et al. 2014. Toxicol Appl Pharmacol. 274:191. PubMed
  134. de Winde CM, et al. 2021. J Cell Sci. 134: . PubMed
  135. Li H, et al. 2015. Reproduction. 150: 417-427. PubMed
  136. Andersen K, et al. 2017. J Am Soc Nephrol. 28:76. PubMed
  137. Oliveira G, et al. 2015. Sci Transl Med. 7: 317ra198. PubMed
  138. Urueña C, et al. 2022. Front Med (Lausanne). 9:991873. PubMed
  139. Reuschl AK, et al. 2022. Cell Rep. 39:110650. PubMed
  140. Chiaro J, et al. 2021. Cancer Immunol Res. 9:981. PubMed
  141. Solleiro-Villavicencio H, et al. 2015. Clin Immunol. . PubMed
  142. Martini E, et al. 2016. Cell Rep. 14:1062-1073. PubMed
  143. Jacobs H, et al. 2016. J Immunol. 196: 3525 - 3531. PubMed
  144. MacDonald K, et al. 2007. Blood . 109:5049. PubMed
  145. Méndez-Lagares G, et al. 2021. J Clin Invest. 131:e148542. PubMed
  146. Mishra P, et al. 2016. PLoS Negl Trop Dis. 10: 0004787. PubMed
  147. Weber A, et al. 2016. Infect Immun . 84: 1413 - 1423. PubMed
  148. Linnerbauer S, et al. 2014. PLoS Pathog. 10:1004068. PubMed
  149. Zheng D, et al. 2022. Front Immunol. 13:808347. PubMed
  150. Kling J, et al. 2011. Exp Parasitol. 129:270. PubMed
  151. Okagawa T, et al. 2016. Infect Immun . 84: 77 - 89. PubMed
  152. Namas R, et al. 2016. Lupus Sci Med. 3: e000148. PubMed
  153. Javed A, et al. 2015. PLoS One. 10: 0142086. PubMed
  154. Li Y, et al. 2015. J Immunol. 195: 1591 - 1598. PubMed
  155. Faustman D, et al. 2012. PLoS One. 7:e41756. PubMed
  156. Alizadeh A, et al. 2018. J Neuroinflammation. 15:53. PubMed
  157. Okano T, et al. 2017. Sci Rep. 7:42412. PubMed
  158. Aggarwal N, et al. 2021. Cell Rep. 37:110170. PubMed

Antigen Details

Cell Type
Tregs
Biology Area
Immunology, 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: 4    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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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

Pricing & Availability
Regulatory Status
RUO
Other Names
FOXP3 Fixation/Permeabilization Buffer
Ave. Rating
Submit a Review
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. No. 424401). Cells were then stained with FOXP3 (clone 206D) Brilliant Violet 421™ (top) or mouse IgG1, κ Brilliant Violet 421™ isotype control (bottom).
Cat # Size Price Quantity Check Availability Save
421403 100 tests NOK1164
Check Availability


Need larger quantities of this item?
Request Bulk Quote
Description

BioLegend's FOXP3 Fix/Perm Buffer Set has been specially formulated for intracellular staining FOXP3 with minimum effect on the surface fluorochrome staining. It contains one 30 ml 4X concentrated FOXP3 Fix/Perm buffer (Cat. No. 421401) and one 25 mL 10X concentrated FOXP3 Perm buffer (Cat. No. 421402). 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. 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.

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.)

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 a 1X working solution with PBS  prior to staining procedures. Dilute 1 part FOXP3 Fix/Perm Buffer (4x) to 3 parts PBS (10 mM sodium phosphate, 150 mM sodium chloride, pH 7.2). 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.

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.
Application References
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  73. Namas R, et al. 2016. Lupus Sci Med. 3: e000148. PubMed
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  76. Faustman D, et al. 2012. PLoS One. 7:e41756. PubMed
  77. Alizadeh A, et al. 2018. J Neuroinflammation. 15:53. PubMed
  78. Okano T, et al. 2017. Sci Rep. 7:42412. PubMed
  79. Aggarwal N, et al. 2021. Cell Rep. 37:110170. PubMed
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  81. Williams JW, et al. 2020. Nat Immunol. 1194:21. PubMed
  82. Danileviciute E, et al. 2022. Nat Metab. 4:589. PubMed
  83. Freimer JW, et al. 2022. Nat Genet. 54:1133. PubMed
  84. Li L, et al. 2022. Cancer Immunol Res. 10:1475. PubMed
  85. Fetit R, et al. 2023. Development. 150:. PubMed
  86. Jin J, et al. 2023. Nat Aging. 3:600. PubMed
  87. Chiu H, et al. 2021. iScience. 24:102748. PubMed
  88. Vella JL, et al. 2021. Life Sci Alliance. 4:. PubMed
  89. Jin J, et al. 2021. Sci Immunol. 6:. PubMed
  90. Hippen KL, et al. 2021. Cytotherapy. 23:704. PubMed
  91. Ren J, et al. 2022. Proc Natl Acad Sci U S A. 119:e2117401119. PubMed
  92. Guttman O, et al. 2022. J Cell Biol. 221: . PubMed
  93. Wang Z, et al. 2022. Proc Natl Acad Sci U S A. 119:e2201899119. PubMed
  94. Zhou Y, et al. 2022. Evid Based Complement Alternat Med. 2022:2197763. PubMed
  95. Hajjar S, et al. 2022. Cell Death Differ. 29:585. PubMed
  96. Yang D, et al. 2016. PLoS Negl Trop Dis. 10: 0004335. PubMed
  97. Singh AK, et al. 2017. JCI Insight. 2:e94275. PubMed
  98. Zbinden A, et al. 2021. Adv Sci (Weinh). 2002500:8. PubMed
  99. Mizraji G, et al. 2017. Cell Rep. 18(2):419-431. PubMed
  100. Puc I, et al. 2022. Int J Mol Sci. 23:. PubMed
  101. Song J, et al. 2022. Clin Transl Med. 12:e992. PubMed
  102. Zemek RM, et al. 2020. Nat Protoc. 15:1628. PubMed
  103. Troutman T, et al. 2012. Proc Natl Acad Sci U S A. 109:273. PubMed
  104. Kawamoto N, et al. 2013. Int Immunol. 25:53. PubMed
  105. Kamigaki T, et al. 2015. Anticancer Res. 35: 4535-4543. PubMed
  106. Fernandez SV, et al. 2020. Breast Cancer Res. 1.009722222. PubMed
  107. Cook K, et al. 2014. Gut. . PubMed
  108. Bamias G, et al. 2007. J Immunol. 178:1809. PubMed
  109. Dang Y, et al. 2007. Clin Cancer Res . 1.932638889. PubMed
  110. Yang Z, et al. 2006. Blood. 107:3639. PubMed
  111. Morandi B, et al. 2013. J Immunol. 191:4858. PubMed
  112. Satoh M, et al. 2016. Sci Rep. 6:28473. PubMed
  113. Zhao XY, et al. 2020. Sci Adv. 6:eaay6191. PubMed
  114. Bankoti R, et al. 2017. Sci Rep. 10.1038/s41598-017-12171-3. PubMed
  115. Basu S, et al. 2015. J Leukoc Biol. 97:279. PubMed
  116. Haller M, et al. 2016. Diabetes. 65(12):3765-3775. PubMed
  117. Van Gool F, et al. 2019. Immunity. 50:362. PubMed
  118. Takamura S, et al. 2010. J Immunol. 184:4696. PubMed
  119. Brennan CA, et al. 2021. Gut Microbes. 13:1987780. PubMed
  120. Scur M, et al. 2022. Nat Commun. 13:7272. PubMed
  121. Greaney SK, et al. 2020. Cancer Immunol Res. 0.504166667. PubMed
  122. Matisz C, et al. 2017. Sci Rep. 7:40631. PubMed
  123. Pfirrmann V, et al. 2015. Cytotherapy. 17: 1139-1151. PubMed
  124. Cecil D, et al. 2014. Cancer Res. 74:2710. PubMed
  125. Liu Y, et al. 2012. Food Chem Toxicol. 50:1920. PubMed
  126. Beavis P, et al. 2011. Proc Natl Acad Sci U S A. 108:16717. PubMed
  127. Zhou Q, et al. 2015. J Immunol. 195: 2493-250. PubMed
  128. Huang S, et al. 2016. Immunol Cell Biol. 10.1038/icb.2016.75. PubMed
  129. Basu S, et al. 2008. J Immunol. 180:5794. PubMed
  130. Luo G, et al. 2021. Sci Rep. 11:7841. PubMed
  131. Keefe RC, et al. 2021. Sci Rep. 11:14933. PubMed
  132. Kwek S, et al. 2015. Cancer Immunol Res. 3: 1008-1016. PubMed
  133. Shiba T, et al. 2014. Toxicol Appl Pharmacol. 274:191. PubMed
  134. de Winde CM, et al. 2021. J Cell Sci. 134: . PubMed
  135. Li H, et al. 2015. Reproduction. 150: 417-427. PubMed
  136. Andersen K, et al. 2017. J Am Soc Nephrol. 28:76. PubMed
  137. Oliveira G, et al. 2015. Sci Transl Med. 7: 317ra198. PubMed
  138. Urueña C, et al. 2022. Front Med (Lausanne). 9:991873. PubMed
  139. Reuschl AK, et al. 2022. Cell Rep. 39:110650. PubMed
  140. Chiaro J, et al. 2021. Cancer Immunol Res. 9:981. PubMed
  141. Solleiro-Villavicencio H, et al. 2015. Clin Immunol. . PubMed
  142. Martini E, et al. 2016. Cell Rep. 14:1062-1073. PubMed
  143. Jacobs H, et al. 2016. J Immunol. 196: 3525 - 3531. PubMed
  144. MacDonald K, et al. 2007. Blood . 109:5049. PubMed
  145. Méndez-Lagares G, et al. 2021. J Clin Invest. 131:e148542. PubMed
  146. Mishra P, et al. 2016. PLoS Negl Trop Dis. 10: 0004787. PubMed
  147. Weber A, et al. 2016. Infect Immun . 84: 1413 - 1423. PubMed
  148. Linnerbauer S, et al. 2014. PLoS Pathog. 10:1004068. PubMed
  149. Zheng D, et al. 2022. Front Immunol. 13:808347. PubMed
  150. Kling J, et al. 2011. Exp Parasitol. 129:270. PubMed
  151. Okagawa T, et al. 2016. Infect Immun . 84: 77 - 89. PubMed
  152. Namas R, et al. 2016. Lupus Sci Med. 3: e000148. PubMed
  153. Javed A, et al. 2015. PLoS One. 10: 0142086. PubMed
  154. Li Y, et al. 2015. J Immunol. 195: 1591 - 1598. PubMed
  155. Faustman D, et al. 2012. PLoS One. 7:e41756. PubMed
  156. Alizadeh A, et al. 2018. J Neuroinflammation. 15:53. PubMed
  157. Okano T, et al. 2017. Sci Rep. 7:42412. PubMed
  158. Aggarwal N, et al. 2021. Cell Rep. 37:110170. PubMed

Antigen Details

Cell Type
Tregs
Biology Area
Immunology, 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: 4    Revision Date: 09.16.2024

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