Cell Stimulation Reagent (with Brefeldin A)

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  • Cell Stimulation Reagent (with Brefeldin A) (500X)
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  • Product Type
    Accessory Reagent
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    BUF077AFdatasheet pdfdatasheet pdf0.1 ml
    BUF077A
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
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    • 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.
    • Intended Use
    • Product Form
      500x concentrate - liquid
    • Reconstitution
    • 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
    • Preservative Stabilisers
    • Purity
    • Approx. Protein Concentrations
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
    • Storage
      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.
    • Shelf Life
      Please see label for expiry date.
    • Acknowledgements
    • Regulatory
      For research purposes only
    • 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 NameYesNoMin DilutionMax 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.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use
      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 (BUF076A) and then add Brefeldin A Solution 1000x (BUF075) or Monensin Solution (BUF074).

    Additional Cell Stimulation Reagent (with Brefeldin A) Formats

    Formats Applications Sizes available
    Cell Stimulation Reagent (with Brefeldin A) : Reagent F 0.1 ml
    • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

    Recommended Secondary Antibody

      Recommended Negative Isotype Control

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Cell Stimulation Reagent (Without Brefeldin A) (500X)BUF076A0.1 mlF
          BUF076A
          Monensin Solution (1000X)BUF0741 mlF
          BUF074
          Brefeldin A Solution (1000X)BUF0751 mlF
          BUF075

          Recommended Positive Controls

            Histology Controls

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