Cells were washed twice in PBS, collected, lysed and boiled in SDS sample buffer at 99°Cfor 5 min. Equal amount of proteins of each cell extracts were resolved by 10–12% Bis-Tris SDS-PAGE and transferred to polyvinylidene difluoride membrane (PVDF, Millipore, Billerica, MA). Non-specific binding was blocked by incubation in 5% skim milk in TBST at room temperature for 1–2 h. Blots were then probed with various primary antibodies, AcH3 (06-599, Millipore, Billerica, MA), H3K9me3 (ab8898, Abcam, Cambridge, UK), Zscan4 (#5114, custom-made), Histone H3 (ab1791, Abcam, Cambridge, UK) and β-actin (P30002, Abmart, Shanghai, China) by overnight incubation at 4°C in 5% skim milk in TBST. Immunoreactive bands were then probed for 1–2 h at room temperature with the appropriate horseradish peroxidase-conjugated secondary anti-Rabbit IgG-HRP (GE Healthcare, NA934V, Piscataway, NJ). The protein bands were detected by Enhanced ECL Amersham™ prime Western blotting detection reagent (GE Healthcare, RPN2232).
β actin p30002
Manufactured by Abmart
Sourced in United Kingdom
β-actin (P30002) is a widely used protein in cell biology research. It is a highly conserved cytoskeletal protein that plays a crucial role in various cellular processes, including cell motility, structure, and division. This protein is commonly used as a reference standard or internal control in various experimental techniques, such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Polyvinylidene fluoride (pvdf), by Merck Group (2 mentions)
Histone h3, by Abcam (1 mentions)
Zscan4, by Abcam (1 mentions)
H3k9me3, by Abcam (1 mentions)
Amersham ecl, by Cytiva (1 mentions)
Chemiluminescent hrp substrate, by Merck Group (1 mentions)
Anti rabbit igg hrp, by GE Healthcare (1 mentions)
2 protocols using β actin p30002
1
Protein Expression and Western Blot Analysis
2
Immunoblotting Analysis of JNK1 Protein
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Cells were washed twice in PBS, collected, and lysed in cell lysis buffer on ice for 30 min and then sonicated for 1 min at 60 of amplitude with 2 s intervals. After centrifugation at 10,000 g, 4°C for 10 min, supernatant was transferred into new tubes. The concentration of the protein sample was measured by bicinchoninic acid, and then protein samples were boiled in SDS Sample Buffer at 99°C for 10 min. 20 μg total proteins of each cell extracts were resolved by 10% Bis-Tris SDS-PAGE and transferred to polyvinylidene difluoride membranes (PVDF; Millipore). Nonspecific binding was blocked by incubation in 5% skim milk in TBST at room temperature for 2 h. Blots were then probed with primary antibodies, JNK1 (SGA0288, Sungene) and β-Actin (P30002, Abmart) by overnight incubation at 4°C in 5% skim milk in TBST. Immunoreactive bands were then probed for 2 h at room temperature with the appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies, anti-Rabbit IgG-HRP (NA934V, GE Healthcare). Protein bands were detected by Chemiluminescent HRP substrate (WBKLS0500, Millipore).
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