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Cell Stimulation Reagent (with Brefeldin A)

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Cell Stimulation Reagent (with Brefeldin A) (500X) thumbnail image 1 Cell Stimulation Reagent (with Brefeldin A) (500X) thumbnail image 2 Cell Stimulation Reagent (with Brefeldin A) (500X) thumbnail image 3 Cell Stimulation Reagent (with Brefeldin A) (500X) thumbnail image 4
Cell Stimulation Reagent (with Brefeldin A) (500X) gallery image 1
Figure A. Unstimulated cells were stained with Alexa Fluor 700 conjugated Mouse anti Human CD3 (MCA463A700) and FITC conjugated Mouse anti Human IFN gamma. Figure B. Cells stimulated with Cell Stimulation Reagent containing Brefeldin A (BUF077A) for 6 hours were stained with Alexa Fluor 700 conjugated Mouse anti Human CD3 (MCA463A700) and FITC conjugated Mouse anti Human IFN gamma (MCA1554F). All experiments performed on red cell lysed human blood gated on lymphoid cells in the presence of 10% human serum. Data acquired on the ZE5 Cell Analyzer.
Cell Stimulation Reagent (with Brefeldin A) (500X) gallery image 2
Figure A. Unstimulated cells were stained with Alexa Fluor 700 conjugated Mouse anti Human CD3 (MCA463A700) and FITC conjugated Rat anti Human IL-2 (MCA1553F). Figure B. Cells stimulated with Cell Stimulation Reagent containing Brefeldin A (BUF077A) for 5 hours were stained with Alexa Fluor 700 conjugated Mouse anti Human CD3 (MCA463A700) and FITC conjugated Rat anti Human IL-2 (MCA1553F). All experiments performed on red cell lysed human blood gated on lymphoid cells in the presence of 10% human serum. Data acquired on the ZE5 Cell Analyzer.
Cell Stimulation Reagent (with Brefeldin A) (500X) gallery image 3
Figure A. Unstimulated cells were stained with Alexa Fluor 700 conjugated Mouse anti Human CD3 (MCA463A700) and FITC conjugated Mouse anti Human TNF alpha. Figure B. Cells stimulated with Cell Stimulation Reagent containing Brefeldin A (BUF077A) for 6 hours were stained with Alexa Fluor 700 conjugated Mouse anti Human CD3 (MCA463A700) and FITC conjugated Mouse anti Human TNF alpha. All experiments performed on red cell lysed human blood gated on lymphoid cells in the presence of 10% human serum. Data acquired on the ZE5 Cell Analyzer
Cell Stimulation Reagent (with Brefeldin A) (500X) gallery image 4
Figure A. Unstimulated cells were stained with FITC conjugated Mouse anti Human CD3 (MCA463F) and purified Rat anti Human TNF alpha (MCA1560) detected with PE conjugated Goat anti Rat (STAR73). Figure B. Cells stimulated with Cell Stimulation Reagent containing Brefeldin A (BUF077A) for 6 hours were stained with FITC conjugated Mouse anti Human CD3 (MCA463F) and purified Rat anti Human TNF alpha (MCA1560) detected with PE conjugated Goat anti Rat (STAR73). All experiments performed on red cell lysed human blood gated on lymphoid cells in the presence of 10% human serum. Data acquired on the ZE5 Cell Analyzer.

Cell Stimulation Reagent (with Brefeldin A) (500X)

Product Type
Accessory Reagent
Specificity
Cell Stimulation Reagent (with Brefeldin A)

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Product Code Applications Pack Size List Price Your Price Qty
BUF077A
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SDS Safety Datasheet SDS
F 0.1 ml loader
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loader

Cell Stimulation Reagent (with Brefeldin A) 500x is a mixture of PMA (phorbol-12-myristate-13-acetate), ionomycin and the protein transport inhibitor Brefeldin A. PMA and ionomycin activate cells and increase cytokine production. Brefeldin A allows for these cytokines to accumulate in the rough endoplasmic reticulum and golgi apparatus allowing for their detection by intracellular fluorescent staining.

Product Form
500x concentrate - liquid
Preparation
500x solution contains: PMA (phorbol-12-myristate 13-acetate) 40.5 μM, Ionomycin 669.3 μM and Brefeldin A 2.5 mg/ml in DMSO
Regulatory
For research purposes only
Guarantee
Guaranteed until date of expiry. Please see product label.

Store at -70oC.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. This product is photosensitive and should be protected from light.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Flow Cytometry
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Instructions For Use
BUF077A
1. Resuspend 1 x106 - 2 x106 cells per ml in cell culture media
2. Thaw the Cell Stimulation Reagent (with Brefeldin A) 500x (BUF077A) in a 37oC water bath
3. To each ml of cell suspension add 2 μl of BUF077A and incubate cells in a CO2, 37oC incubator for 6 hours
4. Harvest cells and centrifuge at 300-400 g for 5 minutes. Discard supernatant
5. To wash the pellet, add 10-20 ml of Cell Staining Buffer (BUF073), vortex to resuspend the pellet then centrifuge at 300-400 g for 5 minutes. Discard supernatant and repeat step 5 for a second time
6. Stain for surface markers or proteins of interest and analyze cells by flow cytometry

Note: Aliquot product to avoid repeat freeze-thawing. Time and culture conditions may need to be optimized.
Activation of cells for >12 hours with Brefeldin A may affect cell viability. In these circumstances it is recommended to use Cell Stimulation Reagent (without Brefeldin A) 500x and then add Brefeldin A Solution 1000x (BUF075) or Monensin Solution (BUF074).

Description Product Code Applications Pack Size List Price Your Price Quantity
Monensin Solution (1000X) BUF074 F 1 ml loader
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Description Monensin Solution (1000X)
Brefeldin A Solution (1000X) BUF075 F 1 ml loader
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Description Brefeldin A Solution (1000X)

BUF077A

157822 158560 161709 162204 163407 164977 165592 165877 166453

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Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"

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