FU97 cell lysates were extracted using RIPA Lysis Buffer containing protease and phosphatase inhibitors (Beyotime), fractionated by SDS‐polyacrylamide gel electrophoresis, and electrotransferred to polyvinylidene difluoride (PVDF) membranes. After blocking in 5% skim milk in triethanolamine buffered saline with 1‰ Tween‐20 (TBST) for 1 h at room temperature, the membranes were incubated with primary antibodies (AFP [1:2500, Abcam, EPR20667], GPC3 [1:1000, HUABIO, EM1709‐60], ALB [1:500, HUABIO, ET1702‐55], E‐cadherin [1:10000, Abcam, EP700Y], N‐cadherin [1:5000, Abcam, EPR1791‐4], VIM [1:1000, Abcam, EPR3776], SNAIL [1:1000, Abcam, EPR21043], and GAPDH [1:5000, Abcam, EPR16891]) at 4°C overnight and then with peroxidase‐conjugated secondary antibodies (HRP Conjugated Goat Anti‐Rabbit IgG [1:5000, HUABIO]) for 1 h at room temperature. After washing thrice with TBST, the blot was soaked in Super ECL Detection Reagent (YEASEN) for 10 s. The membranes were then exposed to a film (Kodak) for 10 s in the dark.
Epr1791 4
Manufactured by Abcam
Sourced in United States
EPR1791-4 is an anti-Arginase-1 antibody produced in rabbit. It is designed for use in Western blotting, immunohistochemistry, and other immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Epr3776, by Abcam (1 mentions)
Ep700y, by Abcam (1 mentions)
Epr16891, by Abcam (1 mentions)
Ripa lysis buffer containing protease and phosphatase inhibitors, by Beyotime (1 mentions)
Hrp conjugated goat anti rabbit igg, by Huabio (1 mentions)
Super ecl detection reagent, by Yeasen (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Enhanced chemiluminescence system, by GE Healthcare (1 mentions)
Sc 271984, by Santa Cruz Biotechnology (1 mentions)
2 protocols using epr1791 4
1
Western Blot Analysis of Liver Cancer Markers
2
Western Blot Analysis of Epithelial-Mesenchymal Transition Markers
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Western blot analysis was performed according to the protocol provided by the manufacturer (Bio-Rad Laboratories Inc., Hercules, CA, USA). Briefly, protein was extracted from cells using radio immunoprecipitation assay (RIPA) lysis buffer (Beyotime, Shanghai, People’s Republic of China). Each sample was separated on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (6%–10% gel) and then transferred onto polyvinylidene difluoride membranes. The polyvinylidene difluoride membranes were blocked with 5% nonfat milk for 2 hours and then incubated with primary rabbit antihuman antibodies for detection of N-cadherin (EPR1791-4 at 1/5,000 dilution; Abcam, San Francisco, CA, USA), E-cadherin (EP700Y at 1/10,000 dilution; Abcam), and ZEB2 (SC-271984 at 1/500 dilution; Santa Cruz Biotechnology Inc., Dallas, TX, USA) overnight at 4°C. The membranes were incubated with a horseradish peroxidase-labeled goat antirabbit IgG antibody for 1 hour. After washing (×4) with Tris-Buffered Saline and Tween 20 Buffer (TBST), the bands were developed using an enhanced chemiluminescence system (GE Healthcare UK Ltd, Little Chalfont, UK). Relative protein expression was normalized to β-actin.
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