|10X Wash Buffer, Phosphate Buffered Saline (PBS) 12528||125 ml|
|4% Formaldehyde, Methanol-Free 47746||20 ml|
|Cell Permeabilization Buffer (Triton™ X-100) 39487||20 ml|
|Flow Cytometry Antibody Dilution Buffer 13616||25 ml|
NOTE: Prior to using this kit, refer to the product webpage for antibodies of interest to determine whether they are validated for Flow Cytometry (F) assays using a Triton X-100 permeabilization protocol.
NOTE: When used in conjunction with fluorescent cellular dyes (including viability dyes, DNA dyes, etc.), please refer to the dye product page for recommended protocol. Visit www.cellsignal.com for a listing of flow cytometry validated cellular dyes.
Additional Reagents (Not Supplied):
NOTE: Not all extracellular antibodies will bind correctly after fixation and/or detergent permeabilization, so a small-scale experiment should be performed if you are unsure. Antibodies targeting CD markers or other extracellular proteins may be added prior to fixation. The antibodies will remain bound to the target of interest during the fixation process. A wash step prior to fixation may be performed but is not necessary.
NOTE: Adherent cells or tissue should be dissociated and in single-cell suspension prior to fixation.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
posted June 2019
revised October 2023
Protocol Id: 1844
The Intracellular Flow Cytometry Kit (Triton X-100) provides the supporting reagents needed to preserve protein states and enable antibodies to bind intracellular targets, for flow cytometric analysis of cells in suspension. Detergent-based permeabilization provides access to the nucleus and other intracellular organelles, without affecting fluorophores of direct conjugates added prior to fixation. This kit contains sufficient reagents for 100 individual samples when following the included protocol.
IMPORTANT: Please refer to the antibody product page to determine if it is validated for use in Flow Cytometry (F) and for information regarding appropriate antibody dilution. Some primary antibodies may require methanol permeabilization, which will be noted on the datasheet. Methanol is not included in this kit.
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