Cell Activation Cocktail (without Brefeldin A)

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Other Names
Cell Stimulation Cocktail, PMA/Ionomycin
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Product Citations
publications
Cell_Activation_Cocktail_without_Brefeldin_1_012815
Human peripheral blood lymphocytes were stimulated with Cell Activation cocktail without Brefeldin A for six hours, then surface stained with CD3 APC and CD154 (clone 24-31) Brilliant Violet 421™ (top) or CD25 (clone BC96) PE (bottom).
  • Cell_Activation_Cocktail_without_Brefeldin_1_012815
    Human peripheral blood lymphocytes were stimulated with Cell Activation cocktail without Brefeldin A for six hours, then surface stained with CD3 APC and CD154 (clone 24-31) Brilliant Violet 421™ (top) or CD25 (clone BC96) PE (bottom).
  • Cell_Activation_Cocktail_without_Brefeldin_2_012815
Cat # Size Price Quantity Avail. Save
423301 100 µl 60€
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423302 400 µl 180€
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Description

Cell Activation Cocktail (without Brefeldin A) is a pre-mixed cocktail with optimized concentration of PMA (phorbol 12-myristate-13-acetate) and ionomycin. Activation of cells by PMA and ionomycin leads to an increase in cell surface expression of certain markers such as CD69 and CD154, which can be detected using this cocktail.

Product Details
Technical Data Sheet (pdf)

Product Details

Preparation
Cell Activation Cocktail (without Brefeldin A) is composed of PMA and ionomycin.
Concentration
Each vial of this cocktail contains phorbol-12-myristate 13-acetate (40.5 µM) and ionomycin (669.3 µM) 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. 423301 contains 100 µL of cocktail, formulated at 500X, that can be used for activating 50 mL of cells of various types.

Cat. No. 423302 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: If investigators are interested in detecting intracellular proteins following activation, this product can be used in conjunction with Monensin (Cat. No. 420701) or Brefeldin A (Cat. No. 420601).

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.

Additional Product Notes

View more applications data for this product in our Scientific Poster Library.

Product Citations
  1. Ayenehdeh J, et al. 2017. Immunology Letters. 10.1016/j.imlet.2017.05.006. PubMed
Publication Library

Antigen Details

Gene ID
NA

Related FAQs

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.

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.

Go To Top Version: 3    Revision Date: 07-03-2019

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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