Western Blot:Article Title: The Ubiquitin E3 Ligase TRIM21 Promotes Hepatocarcinogenesis by Suppressing the p62-Keap1-Nrf2 Antioxidant Pathway.
Article Snippet: The following antibodies were used: AKT (Cell Signaling, Danvers, MA; #4695; 1:1000 for western blotting [WB]), Phospho-AKT308 (Cell Signaling; #2965; 1:1000 for WB), Phospho-AKT473 (Cell Signaling; #4508; 1:1000 for WB), Cytochrome p450 2E1 (Abcam, Cambridge, United Kingdom; ab28146; 1:2000 for WB), Cytochrome p450 2B10 (Santa Cruz, Dallas, TX; sc-53242; 1:2000 for WB), Cytochrome p450 2D1 (1:2000 for WB),55 Cytochrome p450 3A (1:2000 for WB),56 NQO1 (Abcam; ab28947; 1:1000 for WB), Nrf2 (Abcam; ab62352; 1:1000 for WB), GADPH (Cell Signaling; #97166; 1:5000 for WB), b-actin (Cell Signaling; #4970; 1:1000 for WB), b-tubulin (Cell Signaling; #86298; 1:1000 for WB), Lamin B1 (Cell Signaling; #12586, 1:1000 for WB), p62 (Novus Biologicals, Littleton, CO; C11, H00008878MOL; 1:1000 for WB and IF), Keap1 (Proteintech, Rosemont, IL; 10503-2-AP; 1:1000 for WB and IF), Phosphohistone H2A.X (Ser139) (Cell Signaling; #9718; 1:1000 for WB and 1:100 for IHC), Ki-67 (Cell Signaling; #12202; 1:100 for IHC), PCNA (Cell Signaling; #13110; 1:100 for IHC), 8- oxo-dG (R&D Systems, Minneapolis, MN; 4354-MC-050; 1:100 for IHC), cleaved caspase-3 (Cell Signaling; #9661; 1:300 for IHC), HA-Tag (Cell Signaling; #3742; 1:1000 for WB), TRIM21 (Abcam; ab207728; 1:1000 for WB; Proteintech; 12108-1-AP; 1:1000 for WB, 1:100 for IHC), F-4/ 80 (Cell Signaling; #70076; 1:200 for IHC), HSP70 (Abcam; ab2787; 1:200 for IHC).
Article Title: The Ubiquitin E3 Ligase TRIM21 Promotes Hepatocarcinogenesis by Suppressing the p62-Keap1-Nrf2 Antioxidant Pathway
Article Snippet: The following antibodies were used: AKT (Cell Signaling, Danvers, MA; #4695; 1:1000 for western blotting [WB]), Phospho-AKT308 (Cell Signaling; #2965; 1:1000 for WB), Phospho-AKT473 (Cell Signaling; #4508; 1:1000 for WB), Cytochrome p450 2E1 (Abcam, Cambridge, United Kingdom; ab28146; 1:2000 for WB), Cytochrome p450 2B10 (Santa Cruz, Dallas, TX; sc-53242; 1:2000 for WB), Cytochrome p450 2D1 (1:2000 for WB), Cytochrome p450 3A (1:2000 for WB), NQO1 (Abcam; ab28947; 1:1000 for WB), Nrf2 (Abcam; ab62352; 1:1000 for WB), GADPH (Cell Signaling; #97166; 1:5000 for WB), β-actin (Cell Signaling; #4970; 1:1000 for WB), β-tubulin (Cell Signaling; #86298; 1:1000 for WB), Lamin B1 (Cell Signaling; #12586, 1:1000 for WB), p62 (Novus Biologicals, Littleton, CO; C11, H00008878-MOL; 1:1000 for WB and IF), Keap1 (Proteintech, Rosemont, IL; 10503-2-AP; 1:1000 for WB and IF), Phospho-histone H2A.X (Ser139) (Cell Signaling; #9718; 1:1000 for WB and 1:100 for IHC), Ki-67 (Cell Signaling; #12202; 1:100 for IHC), PCNA (Cell Signaling; #13110; 1:100 for IHC), 8-oxo-dG (R&D Systems, Minneapolis, MN; 4354-MC-050; 1:100 for IHC), cleaved caspase-3 (Cell Signaling; #9661; 1:300 for IHC), HA-Tag (Cell Signaling; #3742; 1:1000 for WB), TRIM21 (Abcam; ab207728; 1:1000 for WB; Proteintech; 12108-1-AP; 1:1000 for WB, 1:100 for IHC), F-4/80 (Cell Signaling; #70076; 1:200 for IHC), HSP70 (Abcam; ab2787; 1:200 for IHC).
Immunohistochemistry:Article Title: The Ubiquitin E3 Ligase TRIM21 Promotes Hepatocarcinogenesis by Suppressing the p62-Keap1-Nrf2 Antioxidant Pathway.
Article Snippet: The following antibodies were used: AKT (Cell Signaling, Danvers, MA; #4695; 1:1000 for western blotting [WB]), Phospho-AKT308 (Cell Signaling; #2965; 1:1000 for WB), Phospho-AKT473 (Cell Signaling; #4508; 1:1000 for WB), Cytochrome p450 2E1 (Abcam, Cambridge, United Kingdom; ab28146; 1:2000 for WB), Cytochrome p450 2B10 (Santa Cruz, Dallas, TX; sc-53242; 1:2000 for WB), Cytochrome p450 2D1 (1:2000 for WB),55 Cytochrome p450 3A (1:2000 for WB),56 NQO1 (Abcam; ab28947; 1:1000 for WB), Nrf2 (Abcam; ab62352; 1:1000 for WB), GADPH (Cell Signaling; #97166; 1:5000 for WB), b-actin (Cell Signaling; #4970; 1:1000 for WB), b-tubulin (Cell Signaling; #86298; 1:1000 for WB), Lamin B1 (Cell Signaling; #12586, 1:1000 for WB), p62 (Novus Biologicals, Littleton, CO; C11, H00008878MOL; 1:1000 for WB and IF), Keap1 (Proteintech, Rosemont, IL; 10503-2-AP; 1:1000 for WB and IF), Phosphohistone H2A.X (Ser139) (Cell Signaling; #9718; 1:1000 for WB and 1:100 for IHC), Ki-67 (Cell Signaling; #12202; 1:100 for IHC), PCNA (Cell Signaling; #13110; 1:100 for IHC), 8- oxo-dG (R&D Systems, Minneapolis, MN; 4354-MC-050; 1:100 for IHC), cleaved caspase-3 (Cell Signaling; #9661; 1:300 for IHC), HA-Tag (Cell Signaling; #3742; 1:1000 for WB), TRIM21 (Abcam; ab207728; 1:1000 for WB; Proteintech; 12108-1-AP; 1:1000 for WB, 1:100 for IHC), F-4/ 80 (Cell Signaling; #70076; 1:200 for IHC), HSP70 (Abcam; ab2787; 1:200 for IHC).
Article Title: The Ubiquitin E3 Ligase TRIM21 Promotes Hepatocarcinogenesis by Suppressing the p62-Keap1-Nrf2 Antioxidant Pathway
Article Snippet: The following antibodies were used: AKT (Cell Signaling, Danvers, MA; #4695; 1:1000 for western blotting [WB]), Phospho-AKT308 (Cell Signaling; #2965; 1:1000 for WB), Phospho-AKT473 (Cell Signaling; #4508; 1:1000 for WB), Cytochrome p450 2E1 (Abcam, Cambridge, United Kingdom; ab28146; 1:2000 for WB), Cytochrome p450 2B10 (Santa Cruz, Dallas, TX; sc-53242; 1:2000 for WB), Cytochrome p450 2D1 (1:2000 for WB), Cytochrome p450 3A (1:2000 for WB), NQO1 (Abcam; ab28947; 1:1000 for WB), Nrf2 (Abcam; ab62352; 1:1000 for WB), GADPH (Cell Signaling; #97166; 1:5000 for WB), β-actin (Cell Signaling; #4970; 1:1000 for WB), β-tubulin (Cell Signaling; #86298; 1:1000 for WB), Lamin B1 (Cell Signaling; #12586, 1:1000 for WB), p62 (Novus Biologicals, Littleton, CO; C11, H00008878-MOL; 1:1000 for WB and IF), Keap1 (Proteintech, Rosemont, IL; 10503-2-AP; 1:1000 for WB and IF), Phospho-histone H2A.X (Ser139) (Cell Signaling; #9718; 1:1000 for WB and 1:100 for IHC), Ki-67 (Cell Signaling; #12202; 1:100 for IHC), PCNA (Cell Signaling; #13110; 1:100 for IHC), 8-oxo-dG (R&D Systems, Minneapolis, MN; 4354-MC-050; 1:100 for IHC), cleaved caspase-3 (Cell Signaling; #9661; 1:300 for IHC), HA-Tag (Cell Signaling; #3742; 1:1000 for WB), TRIM21 (Abcam; ab207728; 1:1000 for WB; Proteintech; 12108-1-AP; 1:1000 for WB, 1:100 for IHC), F-4/80 (Cell Signaling; #70076; 1:200 for IHC), HSP70 (Abcam; ab2787; 1:200 for IHC).
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