ubiquitins  (Proteintech)

Journal: Journal of Cancer

Article Title: UBE2C is a diagnosis and therapeutic biomarker involved in immune infiltration of cancers including lung adenocarcinoma

doi: 10.7150/jca.92473

Figure Lengend Snippet: Correlation between cell cycle and apoptosis related genes and UBE2C in LUAD. (A) Correlation between mRNA expression of cell cycle and apoptosis related genes and UBE2C based on GEPIA2 database. (B) Differential protein expression of cell cycle and apoptosis-related genes in LUAD compared with normal lung tissue. (C) After silencing UBE2C, the expression levels of CDK1/2, CASP3, PCNA, MCM4, MCM2, MCM6, MCM7, LMNB2, DFFA, MAP2K2 and TRADD in both NCI-H1299 and A549 cells ( # p >0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Antibodies used in our study included anti-UBE2C (1:2000; ab125002), anti-human β-actin (1:1000, #4970; Cell Signaling Technology, Danvers, MA, USA), anti-p-AKT (1:1000; CST4060), anti-AKT (1:1000; CST4691), anti-p-mTOR (1:1000; CST5536), anti-mTOR (1:1000; CST2972), CDK1/2 (AN21.2, Santa Cruz Biotechnology, sc-53219), PCNA (PC10, Santa Cruz Biotechnology, sc-56), MCM2 (E-8, Santa Cruz Biotechnology, sc-373702), MCM4(G-7, Santa Cruz Biotechnology, sc-28317), MCM6 (H-8, Santa Cruz Biotechnology, sc-393618), MCM7 (141.2, Santa Cruz Biotechnology, sc-9966), CASP3(caspase-3, 31A1067, Santa Cruz Biotechnology, sc-56053), DFFA (LCAD, F-8, Santa Cruz Biotechnology, sc-17816), MAP2K2 (A-1, Santa Cruz Biotechnology, sc-13159), LMNB2 (F-8, Santa Cruz Biotechnology, sc-377379), TRADD (A-5, Santa Cruz Biotechnology, sc-46653) and anti-rabbit IgG (1:1000; #7074, Cell Signaling Technology).

Techniques: Expressing

Journal: Clinical Proteomics

Article Title: Investigation on the mechanisms of human sperm DNA damage based on the proteomics analysis by SWATH-MS

doi: 10.1186/s12014-022-09391-9

Figure Lengend Snippet: Comparisons of differentially expressed proteins and two protein modifications between EXP and CON groups. EXP: The experimental group with sperm DNA fragmentation index (DFI) ≥ 30%; CON: The control group with sperm DFI < 30%; n = 2 for each group. A Western blot showed that the expression levels of RAD23B and DFFA in the experimental group were significantly higher than those in the control group, which were consistent with the results of proteomic analysis. B Sperm proteins were separated by 12% of polyacrylamide gel electrophoresis and stained by Coomassie brilliant blue, and there was no obvious changes in sperm protein levels between the experimental group and control group. C Sperm proteins were separated by 12% of polyacrylamide gel electrophoresis and detected with antibodies against ubiquitins, and the results showed that the ubiquitination modification of sperm proteins at approximately 100 kD in the experimental group was significantly higher than that in the control group. D Sperm proteins were separated by 12% of polyacrylamide gel electrophoresis and detected with antibodies against acetyllysine, and the results showed that the acetylation modification of sperm proteins at 20–45 kD in the experimental group was significantly higher than that in the control group

Article Snippet: Then, the membranes were blocked in tris-buffered saline (TBS) containing 0.1% Tween-20 and 5% nonfat dry milk for 60 min at room temperature, and incubated with antibodies against acetyllysine (PTM-101, 1:1000, PTM BIO, Hangzhou, China), ubiquitins (PTM-1106, 1:1000, PTM BIO, Hangzhou, China), DFFA (ab108924, Abcam, USA) and RAD23B (12,121–1-AP, Proteintech, China) overnight at 4 °C, respectively.

Techniques: Control, Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Staining, Ubiquitin Proteomics, Modification

Journal: Clinical proteomics

Article Title: Investigation on the mechanisms of human sperm DNA damage based on the proteomics analysis by SWATH-MS.

doi: 10.1186/s12014-022-09391-9

Figure Lengend Snippet: Fig. 5 Comparisons of differentially expressed proteins and two protein modifications between EXP and CON groups. EXP: The experimental group with sperm DNA fragmentation index (DFI) ≥ 30%; CON: The control group with sperm DFI < 30%; n = 2 for each group. A Western blot showed that the expression levels of RAD23B and DFFA in the experimental group were significantly higher than those in the control group, which were consistent with the results of proteomic analysis. B Sperm proteins were separated by 12% of polyacrylamide gel electrophoresis and stained by Coomassie brilliant blue, and there was no obvious changes in sperm protein levels between the experimental group and control group. C Sperm proteins were separated by 12% of polyacrylamide gel electrophoresis and detected with antibodies against ubiquitins, and the results showed that the ubiquitination modification of sperm proteins at approximately 100 kD in the experimental group was significantly higher than that in the control group. D Sperm proteins were separated by 12% of polyacrylamide gel electrophoresis and detected with antibodies against acetyllysine, and the results showed that the acetylation modification of sperm proteins at 20–45 kD in the experimental group was significantly higher than that in the control group

Article Snippet: Then, the membranes were blocked in tris-buffered saline (TBS) containing 0.1% Tween-20 and 5% nonfat dry milk for 60 min at room temperature, and incubated with antibodies against acetyllysine (PTM-101, 1:1000, PTM BIO, Hangzhou, China), ubiquitins (PTM-1106, 1:1000, PTM BIO, Hangzhou, China), DFFA (ab108924, Abcam, USA) and RAD23B (12,121–1-AP, Proteintech, China) overnight at 4 °C, respectively.

Techniques: Control, Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Staining, Ubiquitin Proteomics, Modification