Total lysates of primary secretory and maturation enamel cells were prepared in Ripa Buffer (Thermo Fisher Scientific), Protease cocktail inhibitor 100X (Thermo Fisher Scientific), Laemli buffer 4X (BioRad) and β-mercaptoethanol (BioRad) and then loaded at the concentration of 5 μg in 10% SDS-polyacrylamide resolving gels (BioRad). Lysates of HEK-293 cells were prepared as above and used as a negative control. Nitrocellulose membranes were saturated with fat-free milk 5% in TBS (Tris-HCl 50 mM, NaCl 150 mM, pH 7.5) Tween 0.1% for 1 h at room temperature and probed with antibodies against Amelogenin (AMELX, Santa Cruz Biotechnology, sc-32892) and β-Actin (Santa Cruz Biotechnology, sc-47778). Signals were amplified and visualized with horseradish peroxidase-conjugated secondary antibody (Bio-Rad) and enhanced chemiluminescence detected by the Bio-Rad ChemiDoc gel documentation setup. Images of the acquired Western blots were analyzed using the ImageJ software.
Amelx
Manufactured by Santa Cruz Biotechnology
Sourced in United States
AMELX is a protein-coding gene that provides instructions for the production of amelogenin, a major structural protein found in tooth enamel. The product plays a crucial role in the formation and mineralization of tooth enamel.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Bio-Rad (1 mentions)
β mercaptoethanol, by Bio-Rad (1 mentions)
Sds polyacrylamide resolving gels, by Bio-Rad (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Proliferating cell nuclear antigen (pcna), by Abcam (1 mentions)
Hif 1α, by Abcam (1 mentions)
Glut1, by Abcam (1 mentions)
Phosphorylated p53, by Cell Signaling Technology (1 mentions)
2 protocols using amelx
1
Enamel Cell Lysate Western Blot
2
Immunohistochemical Analysis of Dental Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The specimens were embedded in wax using conventional methods. Sections (4 μm thickness) of the specimens were boiled in 10 mM citrate buffer (pH 6.0) for 20 min and cooled at room temperature for 20 min. The specimens were incubated with primary antibodies against HIF-1α (Abcam, CAM, UK; dilution 1:200), phosphorylated p53 (Cell signaling; dilution 1:100), cleaved Caspase-3 (Cell signaling, MA, USA; dilution 1:100), PCNA (Abcam; dilution 1:200), Glut-1(Abcam; dilution 1:200), Glut-2 (Novus biologicals, CR, USA; dilution 1:200), MMP20 (Abcam; dilution 1:200), Amelx (Santa Cruz Biotechnology, TX, USA; dilution 1:200), Dmp1 (LSBio, WA, USA; dilution 1:200), Dspp (Santa Cruz Biotechnology; dilution 1:200) at 4°C overnight. The specimens were incubated with goat anti-rabbit Alexa Fluor 488 (Thermo Fisher Scientific, MA, USA; dilution 1:200), goat anti-mouse Alexa Fluor 555 (Thermo Fisher Scientific; dilution 1:200) antibodies. The sections were counterstained with DAPI (Molecular Probes, OR, USA, dilution 1:1,000) and examined using a confocal laser microscope (DMi8; Leica, Wetzlar, 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!