Proteins extracted from the hepatic cells were separated on 10% SDS-PAGE and transferred to nitrocellulose membranes (Thermo Scientific, USA).The membranes were incubated for overnight at 4 °C with the following primary antibodies: anti-LDLR, -IDOL, -HMGCR, and –LXRα (Abcam, USA), anti-GAPDH (Cell signaling, USA), anti-ABCA1 and -ABCG1 (Novus Biologicals, USA), anti-SREBP1, -SREBP2 (Santa Cruz, USA), anti-HA (BioLegend, USA) and anti-PCSK9 (Proteintech, USA). The blots were then incubated with HRP-conjugated secondary antibody (Thermo Scientific, USA) for 1 h at room temperature and incubated in the Pierce ECL Western Blotting Substrate (Thermo Scientific, USA).
Anti pcsk9
Manufactured by Proteintech
Sourced in United States
Anti-PCSK9 is a laboratory product used to detect and quantify the PCSK9 protein in biological samples. It is designed for research use only and its core function is to provide a tool for researchers to study the PCSK9 protein and its role in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Anti ha, by BioLegend (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
Hrp conjugated beta actin antibody, by Proteintech (1 mentions)
Anti npc1l1, by Novus Biologicals (1 mentions)
Anti ldlr, by Proteintech (1 mentions)
Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 555 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Anti mstn, by Proteintech (1 mentions)
Lsm 880, by Zeiss (1 mentions)
Anti hmgb2, by Proteintech (1 mentions)
Dapi solution, by Wuhan Servicebio Technology (1 mentions)
Anti sod1, by Proteintech (1 mentions)
4 protocols using anti pcsk9
1
Hepatic Protein Expression Analysis
2
Western Blot Analysis of Lipid Regulators
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Cell and liver protein samples were extracted and prepared as previously described [17 (link)]. Proteins were probed using the anti-HMGCR (1 : 1000; Novus), anti-ABCG5 (1 : 500; Proteintech), anti-NPC1L1 (1 : 1000, Novus), anti-PCSK9 (1 : 1000; Proteintech), and anti-LDLR (1 : 1000; Proteintech) and revealed using the horseradish peroxidase (HRP) conjugated secondary antibodies (1 : 2000, Cell Signaling Technology) and HRP-conjugated beta actin antibody (1 : 2000, Proteintech). The housekeeping protein beta-actin was used as an internal reference. Blots were visualized by enhanced chemiluminescence. Densitometry quantification of signals was conducted by ImageJ software.
3
Immunofluorescent Analysis of HK-2 Cells
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After exposure to different stimuli, HK-2 grown on chamber slides were fixed in cold methanol and then incubated with anti-N-tyr and anti-PGC-1α mouse monoclonal antibodies (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at a dilution of 1:100 or with anti-Mstn (dilution 1:100) and anti-PCSK9 (dilution 1:160) rabbit polyclonal antibodies (Proteintech, Manchester, UK). As secondary antibodies, goat-anti-mouse Alexa Fluor 488 or goat-anti-rabbit Alexa Fluor 555 were used (Invitrogen, Carlsbad, CA, USA).
4
Immunofluorescence Analysis of Urogenital Tissues
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Paraffin sections, providing a clear and comprehensive view of the urethra, hindgut, URS, and rectourethral fistulas, were chosen for staining. Subsequently, the sections underwent deparaffinization and antigen retrieval. Blocking was carried out with serum (ZSGB-BIO, Beijing, China) at 25 °C for 1 hour, followed by overnight incubation at 4 °C with primary antibodies. The primary antibodies used were as follows: Anti-Pcsk9 (1:100), anti-Hmgb2 (1:100), and anti-Sod1 (1:100), all sourced from Proteintech (Wuhan, China). After PBS washes, the sections were incubated at 25 °C in the dark for 1 hour with fluorescent secondary antibodies: goat anti-rabbit 488 (1:100; Proteintech). Following PBS washes, the sections were stained with DAPI solution (Servicebio Technology, Wuhan, China) for 10 minutes. Finally, the sections were mounted using an anti-fluorescence fading medium (Solarbio, Beijing, China), and images were captured using a laser-scanning confocal microscope (LSM 880, Zeiss, Oberkochen, Germany).
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