Whole cell lysate were collected using methods previous described (24 (link)). Proteins were separated using a 10% Bis-Tris gel and transferred to an immobilon-P membrane (Millipore). Primary antibodies used included ESR1 (sc-542) (1:1000), PR (sc-7208) (1:1000), and p53 (sc-6243) (1:1000) (Santa Cruz Biotechnology) and signals were detected using the Peirce ECL 2 Western Blotting Substrate (Thermo Scientific).
Esr1 sc 542
Manufactured by Santa Cruz Biotechnology
Sourced in United States
ESR1 (sc-542) is an antibody product offered by Santa Cruz Biotechnology. It is designed for the detection of Estrogen Receptor alpha (ER-alpha) protein expression in various sample types.
Automatically generated - may contain errors
Lab products found in correlation
Immobilon p membrane, by Merck Group (1 mentions)
P53 sc 6243, by Santa Cruz Biotechnology (1 mentions)
Peirce ecl 2 western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Pr sc 7208, by Santa Cruz Biotechnology (1 mentions)
Bradford method, by Bio-Rad (1 mentions)
Hybond ecl, by Cytiva (1 mentions)
Vegfa sc 152, by Santa Cruz Biotechnology (1 mentions)
Imagescanner 3, by GE Healthcare (1 mentions)
Hybond, by Cytiva (1 mentions)
2 protocols using esr1 sc 542
1
Western Blot Analysis of ESR1, PR, and p53
2
Western Blot Quantification of Protein Expression
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Cells were homogenized in RIPA buffer. Protein concentration was determined by Bradford method (Bio-Rad Laboratories, CA). Proteins (25 μg) from cell lysates were separated by electrophoresis, and electrotransferred to nitrocellulose membrane Hybond-ECL (Amersham, Buckinghahmshire, UK). Later, membranes were stained with Ponceau S to be used as the loading control. Membranes were then blocked in TBS-T containing 5% non-fat powdered milk, followed by overnight incubation with primary antibodies (ESR1 Sc-542–1:1000; VEGFA Sc152–1:500; Santa Cruz, Dallas, TX, USA). Blot´s intensity was quantified by densitometry (ImageScanner III, GE Healthcare, Uppsala, Sweden). Results were expressed as arbitrary units in comparison to controls.
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