IO and PO small bowel slides were stained for p-histone 3 (Cell Signaling Technologies; Danvers, MA). The average number of positively stained cells per section was measured in each sample. All slides were analyzed by a single investigator. Quantification of data was performed in GraphPad Prism. Samples from 2-week postoperative C57BL/6 mice were used in both cohorts for proliferation analyses. Western Blot was carried out for p-histone 3 (Cell Signaling Technologies; Danvers, MA). Quantification for Western Blot analysis was performed via Image Lab software (Bio-Rad; Hercules, CA).
P histone 3
Manufactured by Cell Signaling Technology
P-histone 3 is a laboratory reagent used to detect and quantify phosphorylated histone H3, a post-translational modification associated with chromatin structure and cell cycle regulation. It serves as a marker for mitotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Bcl xl, by Cell Signaling Technology (2 mentions)
Phospho bcl 2, by Cell Signaling Technology (1 mentions)
β actin antibody, by ZSGB-BIO (1 mentions)
Phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Enhanced chemiluminescence detection reagent, by Applygen (1 mentions)
Ripa buffer, by Macgene (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Cleavage caspase 3, by Cell Signaling Technology (1 mentions)
Cycb1, by Cell Signaling Technology (1 mentions)
Aurora a, by Cell Signaling Technology (1 mentions)
Mcl 1, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
3 protocols using p histone 3
1
Quantifying Small Bowel Cell Proliferation
2
Western Blot Analysis of Apoptosis Regulators
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Cell pellets were lysed in RIPA buffer (Macgene Technology Ltd.) with 10 μl/ml protease inhibitor cocktail (P8340, Sigma) and 10 μl/ml phosphatase inhibitor cocktail (p0044, Sigma). The proteins in cell lysates were separated by electrophoresis on a 15% SDS–polyacrylamide gel, transferred to nitrocellulose membranes, immunostained, and visualized by enhanced chemiluminescence detection reagents (Applygen Technologies Inc., Beijing, China). Antibodies against Survivin, Bcl-2, Bcl-xL, P-Histone 3, phospho-Survivin, and phospho-Bcl-2 were purchased from Cell Signaling (Danvers, MA), and the antibody specific for phospho-Ser-62-Bcl-xL was purchased from Abcam (Cambridge, UK). The β-Actin antibody was purchased from Zsbio (Beijing, China).
3
Western Blot Analysis of Cell Signaling
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For direct IB analysis, cells were lysed in a Triton X-100 or RIPA buffer with phosphatase inhibitors. The antibodies used were as follows: HA (1:2000; Roche), CycG1 (1:500; Santa Cruz), PARP (1:1000; Cell Signaling), Cleavage-caspase-3 (1:500; Cell Signaling), Aurora A (1:1000; Cell Signaling), NOXA (1:1000; Millipore), Mcl-1 (1:1000; Cell Signaling), Bcl-2 (1:1000; Cell Signaling), Bcl-xL (1:1000; Cell Signaling), CycB1 (1:1000; Cell Signaling), p-Histone 3 (1:500; Cell Signaling) and β-actin (1:5000; Sigma).
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