Protein lysates were obtained from cells using a protein extraction kit (W037, Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Total proteins (30 μg) were subjected to 10% SDS-PAGE electrophoretic separation and the resolved proteins were transferred onto a polyvinylidene difluoride (PVDF) membrane (W016-1-1, Nanjing Jiancheng Bioengineering Institute). Following blocking with non-fat milk powder for 2 h, membranes were incubated with rabbit anti-human SOX8 (1:1000, ab226983, Abcam, Bejing, China), rabbit anti-human cleaved caspase-3 (1:4000, ab49822, Abcam), rabbit anti-human cleaved caspase-9 (1:20,000, ab2324, Abcam), rabbit anti-human β-actin (1:20,000, ab2324, Abcam) at 4°C for 12 h. Subsequently, membranes were incubated with a goat anti-rabbit HRP-conjugated IgG H&L (1:20,000, ab205718, Abcam) secondary antibody for 50 min. The protein bands were visualized using an ECL chemiluminescence kit (32132, ThermoFisher, Shanghai, China) and β-actin was selected as a control to calculate the relative expression of the target proteins.
Ab226983
Manufactured by Abcam
Sourced in China, United Kingdom
Ab226983 is a recombinant monoclonal antibody product manufactured by Abcam. The core function of this product is to bind to a specific target antigen.
Automatically generated - may contain errors
Lab products found in correlation
Ab2324, by Abcam (1 mentions)
Ab205718, by Abcam (1 mentions)
Ab49822, by Abcam (1 mentions)
Ecl chemiluminescence kit, by Thermo Fisher Scientific (1 mentions)
Ab179800, by Abcam (1 mentions)
Ab68183, by Abcam (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Ab231289, by Abcam (1 mentions)
Ab8245, by Abcam (1 mentions)
Ab16048, by Abcam (1 mentions)
2 protocols using ab226983
1
Western Blotting for SOX8 and Caspases
2
Western Blot Analysis of Neuronal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Protein samples were obtained from lysates of primary rat cortical neurons and mouse brain tissue samples using RIPA lysis buffer (Beyotechnology, Shanghai, China). To measure protein expression levels, these samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were then transferred onto polyvinylidene difluoride membranes (Millipore, USA). After blocking with 5% non-fat milk at 4°C for 1 h, the membranes were incubated with anti-cleaved caspase-3 (1:1000, ab231289, Abcam, UK), anti-COX-2 (1:1000, ab179800, Abcam, UK), anti-SOX8 (1:1000, ab226983, Abcam, UK), antiβ-catenin (1:1000, ab68183, Abcam, UK), anti-lamin B1 (1: 1000, ab16048, Abcam, UK), and GAPDH (1:1000, ab8245, Abcam, UK) at 4°C overnight, followed by incubation with the appropriate secondary antibody at room temperature for 1 h. Glyceraldehyde 3-phosphate dehydrogenase was used as the endogenous control. Protein blots were visualized using an ECL chemiluminescence kit (Beyotechnology, Shanghai, China).
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