Freshly isolated MuSCs were plated on tissue culture plates coated with laminin (Roche) and maintained in growth media (45% DMEM, 40% F10, 15% FBS, and 2.5 ng ml−1 basic fibroblast growth factor [bFGF]). Cell proliferation was measured by EdU incorporation (Life Technologies). For clonal analyses, MuSCs were plated individually by limiting dilution in 96-well plates and maintained in growth media for 5 days. Terminal myogenic differentiation was induced with DMEM and 2% horse serum for 3 days.
Edu incorporation
Manufactured by Thermo Fisher Scientific
EdU incorporation is a laboratory technique used to detect and quantify cellular proliferation. It involves the incorporation of the thymidine analog 5-ethynyl-2'-deoxyuridine (EdU) into newly synthesized DNA during the S phase of the cell cycle. This method allows for the identification and analysis of actively dividing cells.
Automatically generated - may contain errors
Lab products found in correlation
Laminin, by Roche (2 mentions)
Alexa fluor 555, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 633, by Thermo Fisher Scientific (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Rabbit anti caspase 3, by Abcam (1 mentions)
Mouse anti pros, by Abcam (1 mentions)
H2ax ser 139, by Merck Group (1 mentions)
Myogenin, by Merck Group (1 mentions)
4 protocols using edu incorporation
1
Isolated Murine Satellite Cell Culture
2
Isolated Murine Satellite Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Freshly isolated MuSCs were plated on tissue culture plates coated with laminin (Roche) and maintained in growth media (45% DMEM, 40% F10, 15% FBS, and 2.5 ng ml−1 basic fibroblast growth factor [bFGF]). Cell proliferation was measured by EdU incorporation (Life Technologies). For clonal analyses, MuSCs were plated individually by limiting dilution in 96-well plates and maintained in growth media for 5 days. Terminal myogenic differentiation was induced with DMEM and 2% horse serum for 3 days.
3
Immunohistochemistry of Drosophila Neural Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Brains were fixed 15 min at 4% paraformaldehyde in 0.1 M HEPES pH 7.4. Antibody staining was performed according to the reported methods (An et al., 2017 (link)). The primary antibodies, dilutions, and sources used in this assay were: rabbit anti-Dan/Danr 1/1000; guinea-pig anti-Dan/Danr 1/1000; mouse anti-Pros 1/10; mouse anti-Elav (44C11) 1/10; rabbit anti-Caspase-3 1/1000 (Abcam); guinea anti-Dpn 1/1000 (Y. Cai’s lab); rabbit anti-Ase 1/1000; rabbit anti-Grh 1/1000 (Y. Cai’s lab); mouse, rabbit and Chicken anti-GFP (Abcam) and rabbit anti-phospho-histone H3 (PH3) (Abcam). Secondary antibodies were conjugated to either Alexa Fluor 488, Alexa Fluor 555, or Alexa Fluor 633 (Molecular Probes), and used at 1/500, 1/1,000, or 1/250, respectively. TO-PRO-3 (Molecular Probes) at 1/5,000 was used for DNA staining and samples were mounted in Vectashield (Vector Laboratories). Images were obtained using OLMPUS upright microscope (FV-1000) and processed in Adobe Photoshop 2021. EdU incorporation was performed as per the kit instructions (Invitrogen).
4
Immunofluorescence Analysis of Cellular Senescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluorescence, BJs and IMR90 fibroblasts were plated on glass coverslips and, when indicated, shifted to DM. Single or double immunofluorescence was performed with the following primary antibodies: MF20 (DSHB), Myogenin (DSHB), Ser139 H2AX (Millipore, 05-636), P16 (Santa Cruz Biotechnology, C-20), MYOD (Dako, 5.8), 53BP1 (Novus, NB100-304), H3S10 (Cell Signaling Technology, 9701S), ABL (Calbiochem, Ab3), and phospho-Y412-ABL (Abcam, ab4717). We used secondary antibodies from Alexa (Life Technologies). DNA synthesis was evaluated by EdU incorporation following the manufacturer's instructions (Invitrogen). Nuclei were visualized by DAPI (4′,6′-diamino-2-phenylindole). The images were analyzed with a LSM5 Pascal Zeiss confocal microscope.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!