According to previously reported method [23 (link)], total proteins were isolated from cells or tissues, separated with sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane. Prior to incubated with antibodies, the membranes were cut and then incubated with anti-rabbit ATG5 antibody (1:1000, #A0203, ABclonal, China), anti-rabbit Beclin-1 antibody (1:1000, #A7353, ABclonal), anti-rabbit transforming growth factor (TGF)-β1 antibody (1:1000, #A7040, ABclonal), anti-rabbit α- smooth muscle actin (SMA) antibody (1:1000, #A17910, ABclonal) and anti-rabbit GAPDH antibody (1:1000, #AC001, ABclonal) overnight at 4 °C. After washing, the membrane was cultured with the second antibody (1:1000, AS029, ABclonal) for 60 min at 37 °C. An enhanced chemiluminescence reagent kit (#K1230, ApexBio, Shanghai, China) was used to visualize the bands.
A0203
Manufactured by ABclonal
Sourced in China
A0203 is a laboratory centrifuge designed for common laboratory applications. It has a maximum speed of 6,000 rpm and a maximum RCF of 3,000 x g. The centrifuge can accommodate a variety of sample tubes and microplates.
Automatically generated - may contain errors
Lab products found in correlation
A7353, by ABclonal (1 mentions)
A7040, by ABclonal (1 mentions)
A17910, by ABclonal (1 mentions)
As029, by ABclonal (1 mentions)
Pvdf membrane, by Beyotime (1 mentions)
Ap0115, by ABclonal (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Beyoecl plus kit, by Beyotime (1 mentions)
2 protocols using a0203
1
Western blot analysis of autophagy and fibrosis
2
Western Blot Analysis of Hepatocyte Proteins
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Hepatocyte samples were treated with RIPA Lysis Buffer (Beyotime, Shanghai, China). Proteins quantified using BCA Protein Assay Kit (Beyotime, Shanghai, China) were separated by SDS-PAGE and transferred onto PVDF membranes (Beyotime, Shanghai, China). The membranes were blocked with 5% skimmed milk/5% BSA in phosphate buffer solution with Tween-20 (PBST) for 1 h and then incubated with antibody against mTOR (AP0115, ABclonal, Boston, MA), SREBP-1c (14088-1-AP, Proteintech, Chicago, IL), LAMP2 (66301-1-Ig, Proteintech), ATG5 (A0203, ABclonal), or β-actin (20657-1-AP, Proteintech) overnight at 4°C. On next day, membranes were probed with an HRP-conjugated secondary antibody (ZSGB-BIO, Beijing, China). Target protein bands were visualized using BeyoECL Plus kit (Beyotime, Shanghai, China), and quantified with ImageJ (NIH, Bethesda, MD).
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