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.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
88-8823	eBioscience Fixation & Permeabilization	N/A	N/A	IC Flow
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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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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