An equal amount of protein samples (40µg) was applied to 4–12% Bis-Tris SDS polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membrane at 30V overnight at 4°C. After blocking with TBS containing 0.05% Tween 20 (TBST) buffer with 5% milk for 1 hour, the blots were probed overnight at 4°C with the following primary antibodies: anti-cholesterol-24-hydroxylase 1A7 (1:500; MAB2259, MilliporeSigma); anti-α-spectrin (1:5000, MAB1622, MilliporeSigma); anti-cleaved caspase-3 (1:1000, # 9664, Cell Signaling Technology Inc.; CST; 3 Trask Lane, Danvers, MA) and anti-β-actin (1:5000, sc-47778, Santa Cruz Biotechnology Inc., Santa Cruz, CA). Appropriate secondary HRP-conjugated antibodies (Santa Cruz) were used. The protein signal was visualized with enhanced chemiluminescence and developed with radiographic film. Image J software was used to measure the mean optical densities (OD) and areas of protein signal on the film after scanning.
Mab2259
Manufactured by Merck Group
MAB2259 is a laboratory equipment product manufactured by Merck Group. It serves as a tool for research and analysis purposes. The core function of this product is to facilitate specific tasks within a laboratory setting. No further details can be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
2 protocols using mab2259
1
Western Blot Analysis of Protein Markers
2
Western Blot Analysis of Protein Markers
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An equal amount of protein samples (40µg) was applied to 4–12% Bis-Tris SDS polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membrane at 30V overnight at 4°C. After blocking with TBS containing 0.05% Tween 20 (TBST) buffer with 5% milk for 1 hour, the blots were probed overnight at 4°C with the following primary antibodies: anti-cholesterol-24-hydroxylase 1A7 (1:500; MAB2259, MilliporeSigma); anti-α-spectrin (1:5000, MAB1622, MilliporeSigma); anti-cleaved caspase-3 (1:1000, # 9664, Cell Signaling Technology Inc.; CST; 3 Trask Lane, Danvers, MA) and anti-β-actin (1:5000, sc-47778, Santa Cruz Biotechnology Inc., Santa Cruz, CA). Appropriate secondary HRP-conjugated antibodies (Santa Cruz) were used. The protein signal was visualized with enhanced chemiluminescence and developed with radiographic film. Image J software was used to measure the mean optical densities (OD) and areas of protein signal on the film after scanning.
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