Sds polyacrylamide resolving gels

Manufactured by Bio-Rad

10% SDS-polyacrylamide resolving gels are laboratory equipment used for the separation and analysis of proteins in a gel electrophoresis setup. These gels are formulated with 10% polyacrylamide and contain sodium dodecyl sulfate (SDS) to denature proteins and provide uniform charge distribution.

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Lab products found in correlation

β mercaptoethanol, by Bio-Rad (2 mentions) Horseradish peroxidase conjugated secondary antibody, by Bio-Rad (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) Laemli buffer, by Bio-Rad (1 mentions) Total oxphos rodent wb antibody cocktail, by Abcam (1 mentions) Chemidoctm xrs, by Bio-Rad (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Image labtm software, by Bio-Rad (1 mentions) Amelx, by Santa Cruz Biotechnology (1 mentions)

2 protocols using sds polyacrylamide resolving gels

Western Blot Analysis of ETC Complexes

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EO total lysates were prepared in Laemli buffer (BioRad) and beta-mercaptoethanol (BioRad) and then loaded in 10% SDS-polyacrylamide resolving gels (BioRad). The 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 ETC complexes (Total OXPHOS rodent WB Antibody Cocktail, Abcam) and beta-Actin (Santa Cruz). Signals were amplified and visualized with horseradish peroxidase-conjugated secondary antibodies (BioRad) and detected by the ChemiDoc^TM XRS + with Image Lab^TM Software (BioRad). Western blots images were analyzed using the ImageJ software (NIH).

Costiniti V., Bomfim G.H., Li Y., Mitaishvili E., Ye Z.W., Zhang J., Townsend D.M., Giacomello M, & Lacruz R.S. (2020). Mitochondrial Function in Enamel Development. Frontiers in Physiology, 11, 538.

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Enamel Cell Lysate Western Blot

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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.

Costiniti V., Bomfim G.H., Neginskaya M., Son G.Y., Mitaishvili E., Giacomello M., Pavlov E, & Lacruz R.S. (2022). Mitochondria modulate ameloblast Ca2+ signaling. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 36(2), e22169.

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