Cell Stimulation Reagent (Without Brefeldin A)

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Cell Stimulation Reagent (Without Brefeldin A) gallery image 1

Figure A. RPE conjugated mouse anti human CD3 (MCA463PE) and FITC conjugated mouse anti human CD69 (MCA2806F) on unstimulated cells. Figure B. RPE conjugated mouse anti human CD3 (MCA463PE) and FITC conjugated mouse anti human CD69 (MCA2806F) on cells incubated with Cell Stimulation Reagent (without Brefeldin A) (BUF076A). All experiments performed on human blood gated on lymphocytes. Data acquired on the ZE5™ Cell Analyzer.

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Figure A. RPE conjugated mouse anti human CD3 (MCA463PE) and FITC conjugated mouse anti human CD154 (MCA1968F) on unstimulated cells. Figure B. RPE conjugated mouse anti human CD3 (MCA463PE) and FITC conjugated mouse anti human CD154 (MCA1968F) on cells incubated with Cell Stimulation Reagent (without Brefeldin A) (BUF076A) for 5 hours. All experiments performed on human blood gated on lymphocytes. Data acquired on the ZE5™ Cell Analyzer.

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  • Cell Stimulation Reagent (Without Brefeldin A) (500X)
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  • Product Type
    Accessory Reagent
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    BUF076AFdatasheet pdfdatasheet pdf0.1 ml
    BUF076A
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
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    • Cell Stimulation Reagent (without Brefeldin A) 500x is a mixture of Phorbol-12-myristate-13-acetate (PMA) and ionomycin. PMA activates protein kinase C and ionomycin acts as a Ca2+ ionophore allowing movement of Ca2+ across membranes. Activation of cells with PMA and ionomycin leads to an increase in surface expression of certain markers such as CD69 and CD154 and the production of cytokines such as IL-2, IFN-gamma and TNF-alpha.
    • Intended Use
    • Product Form
      500x concentrate - liquid
    • Reconstitution
    • Preparation
      500x solution contains: Phorbol-12-myristate-13-acetate (PMA) 40.5 μM and Ionomycin 669.3 μM 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 (without Brefeldin A) 500x (BUF076A) in a 37oC water bath
      3. To each ml of cell suspension add 2 μl of BUF076A 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 using flow cytometry

      Note: Aliquot product to avoid repeat freeze-thawing. Time and culture conditions may need to be optimized. To detect intracellular staining following activation, the Cell Activation Reagent (without Brefeldin A) 500x (BUF076A) can be used with Monensin Solution (BUF074) or Brefeldin A Solution 1000x (BUF075).

    Additional Cell Stimulation Reagent (Without Brefeldin A) Formats

    Formats Applications Sizes available
    Cell Stimulation Reagent (Without 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 (with Brefeldin A) (500X)BUF077A0.1 mlF
          BUF077A
          Monensin Solution (1000X)BUF0741 mlF
          BUF074
          Brefeldin A Solution (1000X)BUF0751 mlF
          BUF075

          Recommended Positive Controls

            Histology Controls

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