DSF-treated HCC cells were subjected to Western blot analysis using anti-p38 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-phospho-p38 (Cell Signaling Technology), and anti-tubulin (Oncogene Science, Cambridge, MA) antibodies. ALDH2-knockdown cells and ALDH1-and ALDH2-double knockdown cells were subjected to Western blotting using anti-ALDH1 (BD Biosciences) and anti-ALDH2 (Abcam, Cambridge, MA) antibodies. GPC3-knockdown cells selected by cell sorting for enhanced green fluorescent protein (EGFP) expression were also subjected to Western blot analysis using anti-GPC3 antibody (Santa Cruz Biotechnology).
Anti aldh2
Manufactured by Abcam
Sourced in United States
Anti-ALDH2 is a primary antibody that recognizes the ALDH2 protein. ALDH2 is an enzyme involved in the metabolism of alcohol and aldehydes. The antibody can be used in various research applications to detect and study the ALDH2 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti 4 hydroxynonenal, by Abcam (2 mentions)
Anti gpc3 antibody, by Santa Cruz Biotechnology (1 mentions)
Anti p38, by Santa Cruz Biotechnology (1 mentions)
Anti phospho p38, by Cell Signaling Technology (1 mentions)
Anti aldh1, by BD (1 mentions)
Anti bcl 2, by Bioss Antibodies (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Anti 4 hne, by Abcam (1 mentions)
Anti lamin b1, by Santa Cruz Biotechnology (1 mentions)
Anti pi3k, by Cell Signaling Technology (1 mentions)
Anti phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
Anti iκbα, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Anti pkcε, by Santa Cruz Biotechnology (1 mentions)
Dynabeads protein a immunoprecipitation kit, by Thermo Fisher Scientific (1 mentions)
Dynabeads protein a, by Thermo Fisher Scientific (1 mentions)
Np40 cell lysis buffer, by Thermo Fisher Scientific (1 mentions)
5 protocols using anti aldh2
1
Western Blot Analysis of Protein Targets
2
Protein Expression Analysis in Lung Tissue
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We isolated the cytoplasmic and nuclear fractions from the lung tissue and determined the protein content using a Bradford assay. Subsequently, lung protein lysates (30 μg per lane) were separated via 10%–12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunoblots were then performed in accordance with the method described previously.9 (link)
The blots were subjected to probing with specific antibodies, including anti-ALDH2, anti-4-HNE (diluted at 1:1000, Abcam, Waltham, MA, USA), anti-PI3K, anti-AKT, anti-pAKT, anti-cleaved caspase 3, anti-NF-κB p65, anti-phospho-NF-κB p65, anti-IκB-α (diluted at 1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-Bcl-2 (diluted at 1:1000, Bioss Inc., Woburn, Massachusetts, USA), anti-lamin B1 (diluted at 1:200, Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (diluted at 1:10,000, Sigma Chemical Company, St. Louis, MO, USA). The data were expressed by determining the relative ratio of the target protein's content to that of the reference protein. The relative ratio of the target protein's content in the control group was established as 1.
The blots were subjected to probing with specific antibodies, including anti-ALDH2, anti-4-HNE (diluted at 1:1000, Abcam, Waltham, MA, USA), anti-PI3K, anti-AKT, anti-pAKT, anti-cleaved caspase 3, anti-NF-κB p65, anti-phospho-NF-κB p65, anti-IκB-α (diluted at 1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-Bcl-2 (diluted at 1:1000, Bioss Inc., Woburn, Massachusetts, USA), anti-lamin B1 (diluted at 1:200, Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (diluted at 1:10,000, Sigma Chemical Company, St. Louis, MO, USA). The data were expressed by determining the relative ratio of the target protein's content to that of the reference protein. The relative ratio of the target protein's content in the control group was established as 1.
3
Co-Immunoprecipitation of ALDH2, 4-HNE, and PKCε
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3T3-L1 preadipocytes were harvested at the indicated times and lysed in NP40 cell lysis buffer (Invitrogen, Carlsbad, CA) for co-immunoprecipitation. Immunoprecipitations were carried out using antibodies coupled to Dynabeads Protein A (Dynabeads® protein A Immunoprecipitation Kit, Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. Immunocomplexes were resolved on 12% SDS-PAGE and then immunoblotted. Antibodies used for immunoprecipitation were anti-ALDH2 (Abcam, Cambridge, MA), anti-4-hydroxynonenal (Abcam, Cambridge, MA) and anti-PKCε (Santa Cruz Biotechnology, Dallas, TX).
4
Western Blot Analysis of Hepatic Proteins
5
Western Blot Analysis of Protein Expression
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Proteins of both mouse tissue and HPASMCs were extracted [35 (link)] and fractionated using 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After that, the proteins were transferred to polyvinylidene difluoride membrane (Millipore) and blocked with 5% bovine serum albumin. Next, the membranes were incubated with the following primary antibodies: β-actin (13000; Wei'ao), rabbit anti-mouse polyclonal antibody, anti-4-hydroxynonenal (Abcam, 11000), anti-ALDH2 (Abcam, 11000), anti-cyclin D1 (CST, 11000), anti-PCNA (CST, 11000), anti-LC3 (CST, 11000), anti-p62 (CST, 11000), anti-ATG5 (Abcam, 11000), anti-ATG7 (Abcam, 11000), anti-Beclin1 (CST, 11000), anti-pPI3K (CST, 11000), anti-PI3K (CST, 11000), anti-pAKT (CST, 11000), anti-AKT (CST, 11000), anti-pmTOR (CST, 11000), anti-mTOR (CST, 11000), and anti-ERK1/2 (CST, 11000). Afterward, the membranes were washed three times using phosphate-buffered saline with Tween-20, and the samples were incubated with anti-mouse or anti-rabbit secondary antibodies (Kangcheng, 13000). Finally, the blots were analyzed using a chemiluminescence detection system (Thermo Fisher Scientific) and quantified using ImageJ gel analysis software.
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