Intracellular Staining Permeabilization Wash Buffer (10X)
Catalog# / Size 421002 / 100 mL
Other Names Perm/Wash, Permeabilization Buffer
Description Intracellular Staining Permeabilization Wash Buffer is useful for intracellular staining
procedures, e.g., in preparation of cells for staining intracellular cytokines or other proteins.
Intracellular Staining Permeabilization Wash Buffer is used to permeabilize cells following
fixation with Intracellular Staining Fixation Buffer (Cat. No. 420801). It is supplied as a 10X
solution and should be diluted in deionized water prior to use. Intracellular Staining
Permeabilization Wash Buffer has been formulated to have minimal effects on cells, reduce
non-specific staining and enhance the signal to noise ratio. It can be used for antibody
dilutions and cell washing during intracellular staining.
Product Details
Storage & Handling The Intracellular Staining Perm wash buffer solution should be stored between 2°C and 8°C. Do not
freeze.
Application ICC, ICFC - Quality tested
IHC - Reported in the literature
Recommended Usage For use in permeabilization, dilute Intracellular Staining Permeabilization Wash Buffer (10X) to 1X in
DI water. Resuspend fixed cells in diluted Intracellular Staining Permeabilization Wash Buffer and
centrifuge at 350 xg for 5-10 minutes, and repeat the process twice. It is recommended that the
reagent be titrated for optimal performance for each application. Please see "Intracellular Cytokine
Staining Protocol" on BioLegend's website for more details.
Application Notes The Intracellular Staining Permeabilization Wash Buffer (10X) may have crystallization or
precipitation observed when it is stored at 2-8°C; however, this is normal and does not affect the
buffer's performance. If there is heavy precipitation observed after dilution to 1X working solution, the
buffer can be filtered to clarify the solution.
Application References
(PubMed link indicates
BioLegend citation)
1. Smeltz RB. 2007. J. Immunol. 178:4786.
2. Kang YJ, et al. 2007. Nature Immunol. 8:601.
3. del Rio ML, et al. 2011. Transplantation. 92:1085. PubMed
4. del Rio ML, et al. 2012. J. Immunol. 188:4885. PubMed
5. Marongiu L, et al. 2013. PLoS One. 8:75684. PubMed
6. Xiao Z, et al. 2013. Mol Immunol. 56:423. PubMed
7. Kusner LL, et al. 2014. PLoS One. 9:102231. PubMed
8. Ni PP, et al. 2014. J Immunol. 193:1778. PubMed
Product Citations
1. Rio M, et al. 2012. J Immunol. 188:4885. PubMed
2. Rio M, et al. 2011. Transplantation. 92:1085. PubMed
3. Marongiu L, et al. 2013. PLoS One. 8:75684. PubMed
4. Xiao Z, et al. 2013. Mol Immunol. 56:423. PubMed
5. Kusner L, et al. 2014. PLoS One. 9:102231. PubMed
6. Ni P, et al. 2014. J Immunol . 193:1778. PubMed
7. Glaser K, et al. 2015. Immunol Lett. 166: 19-27. PubMed
8. Lei F, et al. 2012. J Vis Exp. 63: 3986. PubMed
9. Pearce V, et al. 2015. J Immunol. 195: 3206 - 3217. PubMed
10. Xu Z, et al. 2016. Nat Commun. 7:10728. PubMed
11. Li B, Schmidt N 2016. PLoS One. 11: 0162427. PubMed
12. Ikebuchi R, et al. 2016. Sci Rep. 6:35002. PubMed
Antigen Details
Antigen References 1. Current Protocols in Immunology (John Wiley & Sons New York) Unit 6.24 Detection of Intracellular
Cytokines by Flow Cytometry (Barbara Foster and Calman Prussin NIAID NIH Bethesda MD).
2. Sander B, et al. 1991. Immunol. Rev. 119:65.
3. Sander B, et al. 1993. J. Immunol. Meth. 166:201.
4. Prussin C, et al. 1995. J. Immunol. Meth. 188:117.
Related Protocols
Intracellular Cytokine Staining Protocol - Video
Version: 3 Revision Date: 08/26/2015 
Intracellular Flow Cytometry Staining Protocol
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