Liver tissues from C57BL/6J, Ctrl and miR-181a2b2iECKO mice were homogenized using TissueLyser II (Qiagen) according to manufacturer’ instructions. Proteins were isolated using RIPA buffer (Boston Bio-Productus, BP-115) with protease inhibitor and phosphatase inhibitor. Protein concentrations were determined using Pierce BCA kit (Thermo Scientific). 20 μg protein was loaded per lane on a 4–20% Mini-PROTEAN TGX Gel (Bio-Rad, 456–1096). Separated proteins were transferred to PVDF membranes using the Transfer Turbo Blot system (Bio-Rad) and Trans-Blot Turbo RTA Transfer Kit (Bio-Rad, 170–4272). The membrane was blocked with 5% nonfat milk in TBST for 1 h at room temperature. After blocking, the membrane was incubated overnight at 4 °C with antibodies against PCSK9 (1:1000, ab185194, abcam) or β-actin (1:4000, 4970, Cell Signaling). Quantification of protein bands was performed using a luminescent image analyzer (BioRad, Chemidoc).
Ab185194
Manufactured by Abcam
Sourced in United Kingdom
Ab185194 is a primary antibody produced in rabbit. It is designed for use in immunohistochemistry applications.
Automatically generated - may contain errors
Lab products found in correlation
Tissuelyser 2, by Qiagen (1 mentions)
Pierce bca kit, by Thermo Fisher Scientific (1 mentions)
Chemidoc, by Bio-Rad (1 mentions)
Trans blot turbo rta transfer kit, by Bio-Rad (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Turbo blot transfer system, by Bio-Rad (1 mentions)
Mini protean tgx gel, by Bio-Rad (1 mentions)
Ab192890, by Abcam (1 mentions)
Ab179467, by Abcam (1 mentions)
Ab231137, by Abcam (1 mentions)
E ab 20036, by Elabscience (1 mentions)
3 protocols using ab185194
1
Western Blot Analysis of PCSK9 in Mouse Liver
2
Western Blot Analysis of Autophagy, PCSK9, and SIRT4 Proteins
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Protein lysis, sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE), nitrocellulose membrane transfer, and chemiluminescent acquisition and analysis were conducted as already described [11 (link), 50 (link)] with anti-microtubule-associated proteins 1A/1B light chain 3B (LC3B II/I, 1:2000, ab192890, Abcam, Cambridge, UK), anti-PCSK9 (1:1000, ab185194, Abcam, Cambridge, UK), anti-SIRT4 (1:1000, ab231137, Abcam, Cambridge, UK), anti-α-tubulin (1:5000, E-AB-20036, Elabscience Biotechnology Inc., Houston, TX, USA), and anti-actin (1:3000, ab179467, Abcam, Cambridge, UK) as primary antibodies.
3
Visualizing PCSK9 Expression in Liver
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Immunofluorescence was used to study the expression of PCSK9 in liver sections and cell lines. A rabbit PCSK9-specific antibody (ab185194, Abcam, England and bs-6060r, Bioss, China) was used as the primary antibody. Anti-rabbit IgG (Invitrogen) antibodies were used as the corresponding secondary antibodies. Immunofluorescence images were obtained using a fluorescence microscope (X173; Olympus, Japan).
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