Loading...

Cell Activation Cocktail (with Brefeldin A)

Pricing & Availability
Regulatory Status
RUO
Other Names
Cell Stimulation Cocktail, PMA/Ionomycin
Ave. Rating
Submit a Review
Product Citations
publications
Cell_Activation_Cocktail_with_Brefeldin_1_022814
Human peripheral blood lymphocytes were stimulated with Cell Activation Cocktail (with Brefeldin A) for 6 hours, fixed, permeabilized, and then stained with CD3 APC and TNF-α PE (top) or IFN-γ Brilliant Violet 421™ (bottom).
  • Cell_Activation_Cocktail_with_Brefeldin_1_022814
    Human peripheral blood lymphocytes were stimulated with Cell Activation Cocktail (with Brefeldin A) for 6 hours, fixed, permeabilized, and then stained with CD3 APC and TNF-α PE (top) or IFN-γ Brilliant Violet 421™ (bottom).
  • Cell_Activation_Cocktail_with_Brefeldin_2_022814
Cat # Size Price Quantity Check Availability Save
423303 100 µL 90€
Check Availability


Need larger quantities of this item?
Request Bulk Quote
423304 400 µL 268€
Check Availability


Need larger quantities of this item?
Request Bulk Quote
Description

Cell Activation Cocktail (with Brefeldin A) is a pre-mixed cocktail with optimized concentration of PMA (phorbol 12-myristate-13-acetate), ionomycin, and protein transport inhibitor (Brefeldin A). Activation of cells with PMA and ionomycin leads to cytokine production that is captured in the rough endoplasmic reticulum and Golgi apparatus by the protein transport inhibitor Brefeldin A. Brefeldin A functions to inhibit intracellular protein transport, thus allowing intracellular detection of the secreted cytokine and proteins by immunofluorescent staining.

Product Details
Technical data sheet

Product Details

Formulation
Cell Activation Cocktail (with Brefeldin A) is composed of PMA, ionomycin, and Brefeldin A in DMSO (500X).
Concentration
Each vial of this cocktail contains phorbol-12-myristate 13-acetate (40.5 µM), ionomycin (669.3 µM), and Brefeldin A (2.5 mg/ml) in DMSO (500X).
Storage & Handling
Store cocktail at -70°C upon receipt. Protect from light. Avoid repeat freeze/thaw cycles.
Application

Activ, FC, ICFC - Quality tested
Recommended Usage

Cat. No. 423303 contains 100 µL of cocktail, formulated at 500X, that can be used for activating 50 mL of cells of various types.

Cat. No. 423304 contains 400 µL (4 vials x 100 µL per vial) of cocktail, formulated at 500X, that can be used for activating 200 mL of cells of various types.

Application Notes

Cell Activation Protocol:
1. Resuspend cells of interest in desired cell culture medium. Recommended cell suspension is between 1-2 x 106 cells/mL, although effective activation can be obtained with higher or lower concentration.
2. Completely thaw vials in 37°C water bath.
3. Add 2 µL of the cocktail to each mL of cell suspension.
4. Incubate cells at 37°C in a CO2 incubator for 6 hours or the time period of interest.
5. Harvest activated cells and centrifuge at 350g for 5 minutes. Discard supernatant.
6. Add 2.5 mL of Cell Staining Buffer (Cat. No. 420201) to the cell pellet, mix by vortex or pipetting, then centrifuge at 350g for 5 minutes. Discard supernatant.
7. Repeat step 6.
8. Cells are now ready to be tested for surface markers or proteins of interest.

Note: Activation of cells overnight (>12 hours) with Brefeldin A is generally not recommended due to some level of toxicity. Under these conditions, it is recommended to use Cell Activation Cocktail without Brefeldin A (Cat. No. 423301), and then add monensin (Cat. No. 420701) at appropriate concentrations.

This product is temperature sensitive; repeated freeze/thaw is not recommended.

Intracellular and surface flow staining, as well as ELISA, can be performed following activation of cells with this cocktail.

Product Citations
  1. Benson LN, et al. 2022. Circ Res. 130:1550. PubMed
  2. Chae CS, et al. 2022. Cancer Discov. 12:1904. PubMed
  3. Luo Z, et al. 2022. Int J Mol Sci. 23: . PubMed
  4. Liu Z, et al. 2022. Cancer Commun (Lond). 42:971. PubMed
  5. Hao J, et al. 2022. Cell Rep. 41:111804. PubMed
  6. Mulholland M, et al. 2022. JACC CardioOncol. 4:635. PubMed
  7. Zhao Q, et al. 2023. Nutrients. 15: . PubMed
  8. Wang R, et al. 2023. iScience. 26:105954. PubMed
  9. Sun L, et al. 2023. Mol Med Rep. 27: . PubMed
  10. Sohn J, et al. 2022. Mol Oral Microbiol. 37:42. PubMed
  11. Asrir A, et al. 2022. Cancer Cell. 40:318. PubMed
  12. Shen JZ, et al. 2022. Mol Cell. 82:1123. PubMed
  13. Zhang H, et al. 2022. J Exp Med. 219:. PubMed
  14. Makker P, et al. 2017. PLoS One. 10.1371/journal.pone.0170814. PubMed
  15. Freimer JW, et al. 2022. Nat Genet. 54:1133. PubMed
  16. Li L, et al. 2022. Cancer Immunol Res. 10:1475. PubMed
  17. Jia DJ, et al. 2022. Gut Microbes. 14:2145843. PubMed
  18. Gao L, et al. 2023. J Nanobiotechnology. 21:56. PubMed
  19. Nguyen HM, et al. 2023. Cancers (Basel). 15:. PubMed
  20. Yang Y, et al. 2023. Front Immunol. 14:1033497. PubMed
  21. Schwarz A, et al. 2023. Front Immunol. 14:1038689. PubMed
  22. Yi X, et al. 2023. Signal Transduct Target Ther. 8:107. PubMed
  23. Wu Q, et al. 2023. Theranostics. 13:1381. PubMed
  24. Mohammadpour H, et al. 2023. Cell Rep. 42:112250. PubMed
  25. Zhou C, et al. 2023. Int J Biol Sci. 19:1471. PubMed
  26. Imai H, et al. 2023. iScience. 26:106822. PubMed
  27. Li Q, et al. 2022. Cell Rep. 40:111308. PubMed
  28. Li X, et al. 2022. Nat Commun. 13:2794. PubMed
  29. Volta V, et al. 2021. Nat Commun. 12:6979. PubMed
  30. Zhou T, et al. 2020. Nature. 583:609. PubMed
  31. Pan Y, et al. 2021. Front Cell Neurosci. 15:664312. PubMed
  32. Zhang Y, et al. 2020. Oncol Lett. 1.053472222. PubMed
  33. Jiang Q, et al. 2019. Front Oncol. 1.205555556. PubMed
  34. Rao E, et al. 2021. Sci Immunol. 6:. PubMed
  35. Lin H, et al. 2020. Front Microbiol. 1.330555556. PubMed
  36. Liu F, et al. 2020. Oncoimmunology. 9:1746573. PubMed
  37. Zhou H, et al. 2022. J Clin Invest. 132:. PubMed
  38. Papadas A, et al. 2022. Cell Rep. 40:111201. PubMed
  39. Dai B, et al. 2022. Theranostics. 12:7603. PubMed
  40. Wei JL, et al. 2021. J Immunother Cancer. 9: . PubMed
  41. Liu S, et al. 2017. J Pharmacol Sci. 10.1016/j.jphs.2017.02.001. PubMed
  42. Zongyi Y, et al. 2017. PLoS One. 10.1371/journal.pone.0189617. PubMed
  43. Xu J, et al. 2020. Cell Death Dis. 1.106944444. PubMed
  44. Watkins–Schulz R, et al. 2019. Biomaterials. 205:94. PubMed
  45. Jones KI, et al. 2018. EMBO Mol Med. 10:e9342. PubMed
  46. Du Y, et al. 2022. Nat Commun. 13:231. PubMed
  47. Gudgeon N, et al. 2022. Cell Rep. 40:111193. PubMed
  48. Cho S, et al. 2017. Sci Rep. 7:35. PubMed
  49. Wei W, et al. 2022. mSystems. 7:e0046922. PubMed
  50. Yang J, et al. 2022. Microbiome. 10:149. PubMed
  51. Zhang Y, et al. 2021. J Immunol Res. 2021:5599439. PubMed
  52. Ouyang W, et al. 2021. Invest Ophthalmol Vis Sci. 62:25:00. PubMed
  53. Wang J, et al. 2022. J Clin Invest. :. PubMed
  54. Jain RW, et al. 2018. Cell Rep. 25:3342. PubMed
  55. Lin J, et al. 2021. Nat Commun. 12:5321. PubMed
  56. Weiner, et al. 2021. Nature. 592:450. PubMed
  57. Hao C, et al. 2021. Frontiers in Cell and Developmental Biology. 9:695015. PubMed
  58. Creyns B, et al. 2020. Front Immunol. 1.740972222. PubMed
  59. Sendler M, et al. 2020. Gastroenterology. 158:253. PubMed
  60. Guo Q, et al. 2021. Drug Des Devel Ther. 15:4559. PubMed
  61. Ryan NM, et al. 2022. Front Immunol. 13:932742. PubMed
  62. Pöysti S, et al. 2021. Eur J Immunol. 51:620. PubMed
  63. Koelwyn GJ, et al. 2020. Nat Med. 1452:26. PubMed
  64. Saito E, et al. 2019. Biomaterials. 222:119432. PubMed
  65. Wang Y, et al. 2021. Nat Commun. 12:4964. PubMed
  66. Strobl J, et al. 2021. J Exp Med. 218:. PubMed
  67. Pan S, et al. 2021. Front Cell Dev Biol. 9:629306. PubMed
  68. Mansouri S, et al. 2020. Mucosal Immunol. 0.954861111. PubMed
  69. Gao J, et al. 2022. Front Immunol. 13:955812. PubMed
  70. Deng Z, et al. 2022. Front Oncol. 12:975408. PubMed
  71. Webb J, et al. 2015. Cancer Immunol Res. 3: 926-935. PubMed
  72. Yin S, et al. 2015. Sci Rep. 5: 14432. PubMed
  73. Wu H, et al. 2020. Br J Pharmacol. 177:402. PubMed
  74. Wang X, et al. 2020. Exp Ther Med. 19:3698. PubMed
  75. Li N, et al. 2022. Nat Commun. 13:7281. PubMed
  76. Peng X, et al. 2021. Stem Cell Res Ther. 198:12. PubMed
  77. Chang T, et al. 2020. Aging (Albany NY). 12:21147. PubMed
  78. Li D, et al. 2020. Immunohorizons. 0.661805556. PubMed
  79. Kim ST, et al. 2019. J Immunother Cancer. 0.379166667. PubMed
  80. Liu Y, et al. 2018. Cancer Cell. 33:480. PubMed
  81. Kim ST, et al. 2021. J Immunother Cancer. 9:. PubMed
  82. Yang X, et al. 2021. Bioact Mater. 3150:6. PubMed
  83. Nakao R, et al. 2022. NPJ Vaccines. 7:153. PubMed
  84. Lin J, et al. 2021. Cancer Immunol Res. 9:1413. PubMed
  85. Chen YL, et al. 2022. Front Neurosci. 16:876582. PubMed
  86. Prosser A, et al. 2021. STAR Protoc. 2:100810. PubMed
  87. Liu X, et al. 2021. Adv Sci (Weinh). 8:e2100233. PubMed
  88. Zhang QW, et al. 2021. Am J Cancer Res. 11:2782. PubMed
  89. Xu L, et al. 2022. Front Immunol. 13:841141. PubMed
  90. Wang N, et al. 2020. Front Immunol. 1.765972222. PubMed

Antigen Details

Gene ID
NA

Related FAQs

The Brefeldin A solution freezes at 4°C . Is the solution still OK?

We recommend to thaw in warm water after removing from the fridge.  The Brefeldin A is dissolved in DMSO, which freezes at 18°C, so this is normal.  No one has reported any problems with the freeze/thaw cycles and this is how we recommend it is stored.   If customers are concerned, they can aliquot the Brefeldin A to avoid the freeze thaw.

How do I choose monensin or Brefeldin A solution?

Generally, Brefeldin A is more toxic for longer term incubations, so for shorter stimulations (6 hrs or less) use Brefeldin A.  For longer stimulations use monensin.  We recommend optimization for each cell type and protocol.

Go To Top Version: 3    Revision Date: 07.03.2019

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.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

ProductsHere
Login / Register
Remember me
Forgot your password? Reset password?
Create an Account