Protein lysate of hESCs was prepared in RIPA buffer and NuPAGE® LDS sample loading buffer and denatured for 10 min at 98°C. Samples were run on an SDS- PAGE 10% Bis-Tris gel (ThermoFisher) 60–80 min at 100V, transferred at 100V for 60–70 min, and blocked with 5% non-fat dried milk in 0.1%TBST (0.1% Tween in Tris Buffer Saline solution) for 1 hr. Primary antibody was added to blocking buffer and incubated at room temperature for 1 hr. Secondary antibodies were added after washing with TBST. The PierceTM ECL Western Blotting Substrate (ThermoScientific, 32109) was used on the membrane before film exposure. The primary antibodies used in this study include: rabbit-anti-TFAP2C (Abcam, 76007, 1:1000), mouse-anti-Histone 3 (Abcam, 10799, 1:8000). The secondary antibodies used include: Donkey-anti-goat HRP (Invitrogen, A15999, 1:2000), Sheep-anti-mouse HRP (GE Healthcare Life Sciences, NA931VS, 1:2000).
Sds page 10 bis tris gel
Manufactured by Thermo Fisher Scientific
SDS-PAGE 10% Bis-Tris gel is a laboratory equipment used for the separation and analysis of proteins based on their molecular weight. It is a type of polyacrylamide gel electrophoresis (PAGE) system that utilizes sodium dodecyl sulfate (SDS) to denature proteins and a Bis-Tris buffer system to maintain a consistent pH during the electrophoresis process. The 10% Bis-Tris gel provides an optimal matrix for the separation of a wide range of protein sizes.
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2 protocols using sds page 10 bis tris gel
1
Quantifying TFAP2C and Histone 3 in hESCs
2
Quantifying TFAP2C and Histone 3 in hESCs
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Protein lysate of hESCs was prepared in RIPA buffer and NuPAGE® LDS sample loading buffer and denatured for 10 min at 98°C. Samples were run on an SDS- PAGE 10% Bis-Tris gel (ThermoFisher) 60–80 min at 100V, transferred at 100V for 60–70 min, and blocked with 5% non-fat dried milk in 0.1%TBST (0.1% Tween in Tris Buffer Saline solution) for 1 hr. Primary antibody was added to blocking buffer and incubated at room temperature for 1 hr. Secondary antibodies were added after washing with TBST. The PierceTM ECL Western Blotting Substrate (ThermoScientific, 32109) was used on the membrane before film exposure. The primary antibodies used in this study include: rabbit-anti-TFAP2C (Abcam, 76007, 1:1000), mouse-anti-Histone 3 (Abcam, 10799, 1:8000). The secondary antibodies used include: Donkey-anti-goat HRP (Invitrogen, A15999, 1:2000), Sheep-anti-mouse HRP (GE Healthcare Life Sciences, NA931VS, 1:2000).
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