Product Information
Contents: eBioscience Fixation & Permeabilization Kit
Catalog Number: 88-8823
Sizes: Kit
Storage Conditions: Store Brefeldin A and Permeabilization Buffer at 4° C. Store Fixation Buffer at room temperature.
Handling Conditions: Caution: Harmful if inhaled, swallowed or by contact. Wear proper protective clothing and gloves.
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Questions? Please consult our answers to frequently asked questions at
http://www.ebioscience.com/faq.
Description
The eBioscience Fixation and Permeabilization Kit is designed for use in preparation of living cells for intracellular staining and flow cytometric analysis. A protein transport inhibitor (Brefeldin A) is included for treatment of cells during culture – to cause accumulation of cytokine protein at the Golgi and enhance resulting staining. A fixation buffer is used to cross link, stabilize, and ‘fix’ the cell membrane, while the permeabilization buffer is used to reversibly permeabilize the cell membrane so the staining antibodies can enter the cell effectively.
Components:
- Brefeldin A solution: 1ml. The Brefeldin A Solution is supplied at 1000X working concentration. Prior to addition to the cell cultures, a 1:1000 dilution in the culture media should be made.
- IC Fixation Buffer: 125ml. The IC Fixation Buffer is supplied at 1X working concentration and contains 4% Paraformaldehyde, which is toxic and suspected carcinogen. Contact with eyes, skin and mucous membranes should be avoided.
- Permeabilization Buffer: 100ml. The Permeabilization Buffer is supplied as a 10X concentrate. Prior to use, this should be diluted 10-fold in distilled water. 1X Permeabilization Buffer contains 0.1% saponin (a detergent) and 0.09%sodium azide.
Applications Reported
**For a more detailed protocol of intracellular staining for mouse and human cytokines, see
http://www.ebioscience.com/ebioscience/appls/FCI.htm
References
Sander, B. et al. 1991. Immunol. Rev. 119:65.
Litton, M. et al. 1997. Am. J. Pathol. 150:1607.
Andersson, J. et al. 1994. Immunology 82:16.
Sander, B. et al. 1991. Immunol. Rev. 119:65.
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