The guts were dissected in phosphate-buffered saline (PBS) and fixed organic components with 4% paraformaldehyde for 30 min. Swatches were blocked with special solution for 30 min in a mixture of 0.3% Triton X-100, 0.2% goat serum, and 0.1% fetal calf serum. Samples were incubated in rabbit-derived antibodies against phospho-histone 3 (Millipore, H0412; 1:1500 dilution) overnight at 4 °C, washed three times with PBS with 0.3% Triton X-100, and incubated with anti-rabbit in PBS (Invitrogen, WP20007; 1:1000 dilution) and DAPI dye (Invitrogen, 1:1000 dilution) for 2 h. The treated intestinal samples were observed under a fluorescence microscope (Leica DM4000).
Wp20007
Manufactured by Thermo Fisher Scientific
The WP20007 is a laboratory equipment product produced by Thermo Fisher Scientific. It is designed for general laboratory use. The core function of the WP20007 is to perform a specific task, but a detailed description is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
3 protocols using wp20007
1
Immunohistochemical Analysis of Intestinal Tissue
2
Intestinal Cell Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Ten CR, GF and E. faecium-associated adults (emergence 3 days) were selected, their intestines were dissected in PBS, and fixed in 4% paraformaldehyde solution at room temperature for 30 min. 0.3% PBST (Triton X-100) containing 0.2% goat serum and 0.1% fetal bovine serum was added and incubated for 30 min, rabbit primary antibody PH3 (Millipore, H0412; 1:1000) was added and incubated overnight at 4 °C, and anti-rabbit secondary antibody (Invitrogen, WP20007, 1:1000) was added and incubated for 2 h. PBST was used to wash samples. DAPI (Invitrogen, 1:1000) staining for 10 min, PBST washing, tablet sealing, microscopy image collection (Leica DM4000), and statistical analysis were then performed. Finally, the average value was taken.
3
Intestinal Cell Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ten CR, GF and E. faecium-associated adults (emergence 3 days) were selected, their intestines were dissected in PBS, and fixed in 4% paraformaldehyde solution at room temperature for 30 min. 0.3% PBST (Triton X-100) containing 0.2% goat serum and 0.1% fetal bovine serum was added and incubated for 30 min, rabbit primary antibody PH3 (Millipore, H0412; 1:1000) was added and incubated overnight at 4 °C, and anti-rabbit secondary antibody (Invitrogen, WP20007, 1:1000) was added and incubated for 2 h. PBST was used to wash samples. DAPI (Invitrogen, 1:1000) staining for 10 min, PBST washing, tablet sealing, microscopy image collection (Leica DM4000), and statistical analysis were then performed. Finally, the average value was taken.
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