DPCs were cultured in 24-well plates and stimulated with 100 ng/mL LPS for 24 h. The cells were stained with Hoechst 33342/PI Double Staining Kit (Keygen Biotech Co., Nanjing, China) for 10 min at room temperature and observed under the inverted fluorescence microscope. For fluorescence activated cell sorting (FACS) analysis, cells were digested with EDTA-free trypsin and stained with Hoechst 33342/PI at room temperature for 10 minutes. Blue fluorescent of Hoechst 33342 and red fluorescence of PI were captured with Gallios Flow Cytometer (Beckman coulter, CA, USA).
Hoechst 33342 pi double staining kit
Manufactured by Keygen Biotech
Sourced in China
The Hoechst 33342/PI Double Staining Kit is a laboratory tool used for cell analysis. It contains two fluorescent dyes, Hoechst 33342 and Propidium Iodide (PI), which are used to stain cellular DNA. This kit enables the simultaneous detection and differentiation of live and dead cells in a sample.
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2 protocols using hoechst 33342 pi double staining kit
1
Assessing Cell Viability via Hoechst/PI Staining
2
Evaluating Pyroptosis in Mouse Osteoblasts
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The pyroptosis of mouse osteoblasts was evaluated using the lactate dehydrogenase (LDH) release kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Cell culture supernatants were collected in order to assay the cells for lactate dehydrogenase activity to assess the integrity of the cell membrane. An enzyme marker used in spectrophotometry had its absorbance measured at 450 nm. LDH activity was calculated by the formula = (OD of assay group - OD of control group)/(OD of standard group - OD of blank group) × concentration of standard × N × 1000.
Staining of mouse osteoblasts with Hoechst 33342/PI double staining kit (KeyGEN BioTECH, Jiangsu, China). Following walling, mouse osteoblasts were transplanted onto 24-well plates at a density of 1 × 104 cells per well. The cells were then double stained with Hoechst 33342/PI. 3 μL of Hoechst 33342 was used to stain the cells for 10 min at 37 °C, then with 3 μL PI at room temperature for 10 min before being examined under a fluorescence microscope.
Staining of mouse osteoblasts with Hoechst 33342/PI double staining kit (KeyGEN BioTECH, Jiangsu, China). Following walling, mouse osteoblasts were transplanted onto 24-well plates at a density of 1 × 104 cells per well. The cells were then double stained with Hoechst 33342/PI. 3 μL of Hoechst 33342 was used to stain the cells for 10 min at 37 °C, then with 3 μL PI at room temperature for 10 min before being examined under a fluorescence microscope.
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