Cell lysates containing ~30 μg protein were loaded on a 15-well 4 to 12% NuPage gel and electrophoresed for 45 min at 150V. Proteins were transferred to nitrocellulose using the Transblot Turbo (BioRad). Blots were rinsed with water and dried, reconstituted with TBS, and blocked with Odyssey TBS blocking buffer for 1 h. Antibodies were added at 1:1000 dilution (anti-Adpgk, Abcam, ab228633) or 1:5000 dilution (anti-actin, Cell Signaling Technology, 3700) in TBS blocking buffer, 0.2% Tween-20 and rocked overnight at 4 °C. Blots were then rinsed four times with TBS, 0.1% TBST. Secondary antibodies were added at 1:15,000 dilution (LI-COR, goat anti-mouse 680, goat anti-rabbit 800) for 1 h at room temperature. Blots were rinsed three times with TBS, 0.1% TBST, then once with TBS. Images were obtained using an Odyssey imager (LI-COR).
Ab228633
Manufactured by Cell Signaling Technology
Ab228633 is a laboratory equipment product manufactured by Cell Signaling Technology. The core function of this product is to serve as a tool for scientific research and analysis. Further details about the specific applications or intended use of this product are not available.
Automatically generated - may contain errors
2 protocols using ab228633
1
Western Blot Analysis of Adpgk and Actin
2
Western Blot Protein Detection Protocol
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Cell lysates containing ~30 µg protein were loaded on a 15-well 4-12% NuPage gel and electrophoresed for 45 min at 150V. Proteins were transferred to nitrocellulose using the Transblot Turbo (BioRad). Blots were rinsed with water and dried, reconstituted with TBS, and blocked with Odyssey TBS blocking buffer for 1 h. Antibodies were added at 1:1000 dilution (anti-Adpgk, Abcam, ab228633) or 1:5000 dilution (anti-actin, Cell Signaling Technology, 3700) in TBS blocking buffer, 0.2% Tween-20, and rocked overnight at 4°C. Blots were then rinsed four times with TBS, 0.1% TBST. Secondary antibodies were added at 1:15000 dilution (LI-COR, goat anti-mouse 680, goat anti-rabbit 800) for 1 h at room temperature. Blots were rinsed three times with TBS, 0.1% TBST, then once with TBS. Images were obtained using an Odyssey imager (LI-COR).
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