Cleaved PARP (Asp 214) (F21-852)
Select Conjugate
About Cleaved PARP (Asp 214)
ISOTYPE
IgG1, k
CYTOMETER USED
LSRII
EXPERIMENT CELL SOURCE
Proliferating PBMC
REACTIVE SPECIES
Human
HOST SPECIES
Mouse
cPARP (Asp 214) FITC
| Perm I | Perm II | Perm III | Perm IV (0.5x) | Perm IV (1.0x) | |
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| Intact Cells |
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| Ungated |
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cPARP (Asp 214) PE
| Perm I | Perm II | Perm III | Perm IV (0.5x) | Perm IV (1.0x) | |
|---|---|---|---|---|---|
| Intact Cells |
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| Ungated |
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cPARP (Asp 214) A647
| Perm I | Perm II | Perm III | Perm IV (0.5x) | Perm IV (1.0x) | |
|---|---|---|---|---|---|
| Intact Cells |
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| Ungated |
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cPARP (Asp 214) A700
| Perm I | Perm II | Perm III | Perm IV (0.5x) | Perm IV (1.0x) | |
|---|---|---|---|---|---|
| Intact Cells |
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| Ungated |
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Gating Hierarchy
Intact Cells
Intact Cells
Protocol & Other Details
T cells from human PBMC were induced to proliferate using beads coated with anti-CD3 and anti-CD28 (Invitrogen, Cat. No. 111.31D). After 72 hours in culture, proliferating cells were treated with 10 μM camptothecin (Sigma, Cat. No. C-9911) for 5 hours at 37°C to induce apoptosis.
In comparison with other permeabilization conditions, a substantially higher percentage of cells stained positive for cleaved PARP when Perm Buffer IV (1X) was used. Therefore, the Perm Buffer IV (1X) permeabilization is not recommended for cleaved PARP staining.
In comparison with other permeabilization conditions, a substantially higher percentage of cells stained positive for cleaved PARP when Perm Buffer IV (1X) was used. Therefore, the Perm Buffer IV (1X) permeabilization is not recommended for cleaved PARP staining.
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