Cells were grown to confluence (80%) on coverslips coated with poly-lysine (Sigma, USA), followed by treatment with either metformin or compound C for indicated time points, and were washed twice with 1X PBS prior to fixation in 4% formalin for 10 min at room temperature. Cells were washed twice in 1X PBS. Prior to blocking, cells were permeabilized in 0.2% TRITON-X-100 (Sigma, USA) for 15 min followed by washes with 1X PBS. Bocking was done in 5% bovine serum (Hi-media, India) for 1 h at room temperature. Cells were stained with RUNX1/AML1 and AMPK or STAT3 for 2 h at room temperature, followed by the respective fluorescence-tagged secondary antibody staining for 1 h (Alexa Fluor 488 and 594, Invitrogen, USA). Cells were counterstained with 4′,6-diamino-2-phenylindole (DAPI) (Thermo Scientific, USA) for nuclei and images were captured using a laser scanning confocal microscope (LSM 780, Carl Zeiss, Germany).
Bovine serum
Manufactured by HiMedia
Sourced in India
Bovine serum is a cell culture media supplement derived from the blood of cattle. It provides a source of proteins, growth factors, and other essential nutrients required for the growth and maintenance of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780, by Zeiss (2 mentions)
Polylysine, by Merck Group (1 mentions)
Alexa fluor 488 and 594, by Thermo Fisher Scientific (1 mentions)
4 6 diamino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Anti oct4, by BioLegend (1 mentions)
Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
2 protocols using bovine serum
1
RUNX1/AMPK Colocalization Microscopy Protocol
2
Immunofluorescence Analysis of hUMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Upscaled hUMSCs at passage 3 were seeded on cover slip. Next they were fixed in 4% paraformaldehyde for 15 min at 4 °C followed by permeabilization in 0.2% Triton X-100. After blocking in bovine serum (HiMedia Laboratories Pvt. Ltd., India), the cells were washed gently. Primary antibodies, anti-Oct-4 (Biolegend), and FITC mouse anti-human CD90 (Biolegend) were incubated with cells for 2 h. After washing with PBS solution, a secondary antibody, namely, Alexa fluor 488 goat anti mouse IgG (Invitrogen, Carlsbad, CA, United States), was added. Cover slip was mounted, and fluorescent images were captured using spinning confocal microscope (Zeiss, Germany).
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!