Brain tissues were removed, fixed, and rinsed with 0.1 M PBS solution until the brains exhibited no irritating odor. The brains were sliced sagitally in a mold at a thickness of 4 μm, placed in a tissue embedding box, and soaked in 75% alcohol for 24 h. After dehydration in gradient ethanol solutions, the slices were placed at 55°C and dried. The slices were placed in xylene I for 5 min followed by xylene II for 5 min and then hydrated in gradient alcohol solutions. Proteinase K without DNase was added dropwise to the slices, and the slices were incubated in a 37°C incubator for 30 min and rinsed with PBS. TUNEL reaction solution (G3250, PROMEGA) was added dropwise to the slices, and the slices were incubated in a 37°C incubator in the dark for 1 h, rinsed with PBS, dried, and mounted. TUNEL staining was observed with a digital pathological section (fluorescence) scanning analyzer. The green FIOD values of TUNEL in the same area of the cerebral cortex were measured using Image-Pro Plus software 6.0 and statistically analyzed (FIOD/area∗100).
Tunel reaction solution g3250
Manufactured by Promega
The TUNEL reaction solution (G3250) is a key component for the TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) assay, which is used to detect and quantify apoptosis or programmed cell death. The solution contains the necessary reagents, including terminal deoxynucleotidyl transferase and labeled nucleotides, to facilitate the TUNEL reaction.
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Lab products found in correlation
2 protocols using tunel reaction solution g3250
1
Quantitative TUNEL Staining Analysis
2
TUNEL Assay for Apoptosis in Rat Hippocampus
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On day 6, the rats were deeply anesthetized and perfused with PBS until the brains exhibited no irritating odor. The brains were then cut into 4 µm-thick coronal slices in a mold, placed in a tissue embedding box, and soaked in 75% alcohol for 24 h. Next, the slices were dehydrated in gradient ethanol solutions at 55 • C and dried. The slices were then placed in xylene I for 5 min followed by xylene II for 5 min and hydrated in gradient alcohol solutions. The slices were covered with Proteinase K without DNase (provided by a kit), incubated in a 37 • C incubator for 30 min and washed with PBS. TUNEL reaction solution (G3250, PROMEGA) was added dropwise to the slices, which were then incubated at 37 • C in the dark for 1 h, washed with PBS, dried, and mounted. A digital pathological section (fluorescence) scanning analyzer (OLYMPUS, Tokyo, Japan) was used to assess the TUNEL staining results. The green FIOD values of TUNEL in the same area of the right hippocampus were determined using Image-Pro Plus software 6.0 and statistically analyzed (FIOD/area × 100).
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