This was performed following standard protocols [43] (link). Briefly, 5µg of LDL and HDL samples were resolved on 5% SDS-PAGE gel and transferred to PVDF membrane. Membrane was blocked in 5% bovine serum albumin (BSA) and incubated with apoB antibody specific to amino-terminal human apoB peptide (Santa Cruz Biotechnologies) followed by peroxidase-conjugated secondary antibody. Specific bans were detected by measuring chemiluminescence using an ECL Advanced Western Blotting Kit (GE Healthcare, Piscataway, NJ).
Ecl advanced western blotting kit
Manufactured by GE Healthcare
Sourced in United States
The ECL Advanced Western Blotting Kit is a laboratory equipment designed for the detection and analysis of proteins in Western blot applications. It provides a sensitive and efficient chemiluminescent detection system for the visualization of target proteins.
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2 protocols using ecl advanced western blotting kit
1
Western Blot Analysis of Apolipoproteins
2
Western Blot Analysis of Hsp90 Expression
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Cell lysates were resuspended to equal total protein concentrations as determined by a NanoDrop 2000 spectrophotometer (Thermo Fischer Scientific, Inc.) Proteins resolved by SDS-PAGE gels were transferred onto Hybond Supported Nitrocellulose membranes (GE Healthcare Life Sciences, Chalfont, UK) membrane. Hsp90 was detected by chemiluminescence using the ECL Advanced Western Blotting kit (GE Healthcare Life Sciences), after incubation with a horseradish peroxidise-conjugated donkey anti-mouse secondary antibody, using the Chemidoc™ EQ system (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Images were analysed using ImageJ software. Anti-Histone H3 (#9715; used at 1:2,500 dilution) and anti-Hsp90α/β (sc-13119; used at 1:1,000 dilution) antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA) and Santa Cruz Biotechnology, Inc., respectively.
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