Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
New Zealand (English)
Profile Icon Profile Icon
Sign-in/Register UserName
FACSelect

Link Arrow
nand

Link Arrow
test rollout page

Link Arrow
Solutions

Link Arrow
Discover & Learn

Link Arrow
Resources & Tools

Link Arrow
Support

Link Arrow
test-page-don't-publish

Link Arrow
TestPage

Link Arrow
Products

Link Arrow
Close Menu
Alert Icon
Gating Hierarchy
Intact Cells
Singlets Singlets
Intact Cells Intact Cells
Protocol & Other Details
Following fixation and permeabilization, cells were stained with 7-AAD for 15 minutes at room temperature. http://www.bdbiosciences.com/documents/BD_FlowCytometry_DNA_Staining_Protocol.pdf
Notes
Human Surface Marker Antibody Concentrations:
Antibody titration is critical for successful post-permeabilization staining of CD markers and other cell surface antigens. The 1X concentration represents cells stained with the recommended test size for live cell stains (see product Technical Data Sheets for test size information). The 1/4X or 1/16X concentrations represent cells stained with 1/4 or 1/16 the antibody recommended for live cell stains.
Mouse Surface Marker Antibody Concentrations:
Antibody titration is critical for successful post-permeabilization staining of CD markers and other cell surface antigens. The 1X concentration represents cells stained with the recommended test size for live cell stains. The 1/4X or 1/16X concentrations represent cells stained with 1/4 or 1/16 the antibody recommended for live cell stains.
YG PE, YG PE-Cy7, and YG PE-Cy5:
PE, PE-Cy5, and PE-Cy7 fluorochromes may be excited by either a 488-nm blue laser or a 561-nm yellow-green laser. Since excitation by the yellow-green laser improves PE detection sensitivity, data collected from the 561-nm laser detection array (YG PE, YG PE-Cy5, and YG PE-Cy7) are provided along with data from the 488-nm laser detection array (PE, PE-Cy5, and PE-Cy7) when available.
Fold-Change Statistic:
Fold-change of a channel (e.g. pStat5-PE) is calculated as the ratio between the median fluorescence intensity of the stimulated sample (e.g. IL-2) and the median fluorescence intensity of the unstimulated sample (e.g. Control).
Percentile Distance Statistic:
Percentile distance is a measure of the spread between the 95th percentile and the 5th percentile of the fluorescence intensity of a channel (e.g. CD3-FITC). The distance is calculated as the ratio between the 95th percentile of a channel and the 5th percentile of a channel in its transformed space (log, linear, arcsinh or biexponential).
Permeabilization Buffers
BD Phosflow™ Perm/Wash Buffer I (557885) BD Phosflow™ Perm Buffer II (558052) BD Phosflow™ Perm Buffer III (558050) BD Phosflow™ Perm Buffer IV (560746)

Perm Buffer IV 1X vs. 0.5X:

 

Perm Buffer IV (provided as a 10X stock solution) may be used at a 1X or 0.5X concentration. The higher (1X) concentration provides optimal resolution of certain intracellular phosphoprotein stains but might result in increased cell loss and decreased ability to stain certain cell surface markers.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Alexa Fluor, Pacific Blue and Texas Red are trademarks of Life Technologies Corporation. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva. AmCyan is a trademark of Takarabio Inc. BD-40149 (v1.0) 0721

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.