Senescence Cells Histochemical Staining Kit

Protocol tips

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	Add 1 ml of the Staining Mixture per well and incubate at 37 °C without CO2 until the cells are stained blue

Protocol tips
Add 1 ml of the Staining Mixture per well and incubate at 37 °C without CO2 until the cells are stained blue

Publication protocol

hMSC and hMSC TERT cells were passage into two T flasks each at 5000 cells/cm without hyg for 1 week and then harvested and counted. Similar cultures were combined and passaged again into two T flasks. Using cumulative counts, the population-doubling level (PDL) was calculated at each passage as follows: PDL=log (/)/log 2, where =passage number, =final number of cells, and =number of cells seeded at passage. After each passage, PDL was added to the previous PDL to get the new PDL. Serial passage levels were expressed as PDL or as the passage number as P(), where is the passage number after transduction; and is the passage number from the original primary culture. At each passage, samples were taken for RNA analysis using the Qiagen RNeasy Miniprep kit (Qiagen, Valencia, CA) and telomerase activity using the TeloTAGGG telomerase polymerase chain reaction (PCR) ELISA (Roche Applied Science, Indianapolis, IN). Cell proliferation was studied using the Cell Proliferation ELISA and BrdU (colometric) Assay (Roche Applied Science); cell senescence using the Senescence Cells Histochemical Staining Kit (Sigma-Aldrich); and apoptosis using the TiterTACS™ apoptosis detection kit from R&D Systems, Inc. (Minneapolis, MN).

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Download the product protocol from Sigma-Aldrich for Senescence Cells Histochemical Staining Kit below.

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