Immunostaining was performed on 5-μm paraffin-embedded sections from the dorsal skin. The sections were deparaffinized, rehydrated, and boiled in citrate buffer solution. After blocking in 10% goat serum (in PBS), the sections were incubated with the following primary antibodies: mouse monoclonal anti-β-catenin (1:100, Santa Cruz, USA, sc-7963), rabbit polyclonal anti-Wnt10b (1:100, Santa Cruz, USA, sc-25524), mouse monoclonal anti-BrdU (1:200, Abcam, USA, ab8039), rabbit polyclonal anti-Sox4 (1:100, Santa Cruz, USA, sc-20090), mouse monoclonal anti-β1-integrin (1:100, Santa Cruz, USA, sc-9970), and rabbit polyclonal anti-CD34 (1:100, Santa Cruz, USA, sc-9095). CY3-labeled secondary antibodies (goat anti-rabbit, A0516 and goat anti-mouse, A0521, Beyotime, China) were used for immunofluorescence, and DAPI (Beyotime, C1002) was used to counterstain the cell nuclei. An HRP-labeled goat anti-mouse detection kit (Zhongshan Goldenbridge, China, PV-6002) was used for immunohistochemistry.
Mouse monoclonal anti brdu
Manufactured by Abcam
Sourced in United States
Mouse monoclonal anti-BrdU is a primary antibody that recognizes 5-bromo-2'-deoxyuridine (BrdU), a synthetic thymidine analog. It can be used to detect and quantify cell proliferation in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti wnt10b, by Santa Cruz Biotechnology (1 mentions)
Anti sox4, by Santa Cruz Biotechnology (1 mentions)
Mouse monoclonal anti β catenin, by Santa Cruz Biotechnology (1 mentions)
A0521, by Beyotime (1 mentions)
Anti cd34, by Santa Cruz Biotechnology (1 mentions)
C1002, by Beyotime (1 mentions)
A0516, by Beyotime (1 mentions)
3 protocols using mouse monoclonal anti brdu
1
Immunostaining of Skin Tissue
2
Visualizing ssDNA with BrdU Immunolabeling
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To visualize ssDNA, cells were cultured with BrdU either 11 h prior to, and during, CPT treatment, or during CPT treatment only. No difference in overlap quantification was seen between approaches. After cell fixation and the naDNA “Click” reaction, BrdU was then immunolabeled alongside protein with mouse monoclonal anti-BrdU (Abcam, 8039). This approach, without any denaturation of the DNA, has previously been demonstrated as limiting antibody access to, and fluorescent tagging of, the ssDNA only30 (link). Proteins of interest were labeled using either direct or indirect labeling with Alexa Fluor 488 and 568 fluorophore-labeled antibodies that have either been previously validated in IF experiments or were cross-validated by us as listed in Supplementary Table 4 .
3
Quantifying Cardiac Fibroblast Proliferation
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Bromodeoxyuridine (BrdU) staining was used to assess the proliferation of cardiac fibroblasts cultured on CNH-Col and collagen substrates. On days 1, 3, and 7, BrdU reagents (20 μmol/ml) were incorporated into the medium of cardiac fibroblasts and incubated for 4 h. After fixing, the cardiac fibroblasts were denaturized with hydrochloric acid (2 mol/l) and then neutralized with sodium borate (0.1 mol/l) for 12 min at room temperature. Mouse monoclonal anti-BrdU (1:100, Abcam) was used to stain proliferative cells at 4 °C overnight. Alexa Fluor 548-conjugated secondary antibodies (1:500, Invitrogen) were utilized to conjugate with the primary antibody. Then, the cells were counterstained with DAPI in D-PBS. Finally, the cells were visualized under a fluorescence microscope in 10 randomly selected fields for each group. The images were analyzed by ImageJ software and Image-Pro Plus 6.0.
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