Trim21

Manufactured by Cell Signaling Technology

Sourced in United States

TRIM21 is a protein that functions as an E3 ubiquitin ligase. It is involved in the ubiquitination and subsequent proteasomal degradation of target proteins.

Automatically generated - may contain errors

Lab products found in correlation

Phospho akt, by R&D Systems (1 mentions) Anti hur, by Santa Cruz Biotechnology (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Ubiquitin, by Cell Signaling Technology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Igf2bp3, by Abcam (1 mentions) Hrp labeled rabbit igg, by Cell Signaling Technology (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Hnrnpa1, by Cell Signaling Technology (1 mentions) Cox 4, by Cell Signaling Technology (1 mentions) Sodium formate, by Merck Group (1 mentions) Sirt3, by Cell Signaling Technology (1 mentions) Tublin, by Beyotime (1 mentions) Sab4301135, by Abcam (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Mg132, by Merck Group (1 mentions) Acetylated lysine, by Cell Signaling Technology (1 mentions) Nh4cl, by Sangon (1 mentions)

4 protocols using trim21

Western Blot Analysis of MCF7 Cell Lysates

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Lysates of MCF7 cells in S10 lysis buffer were resolved by 10% or 12% SDS-PAGE, transferred to PVDF membrane (MilliporeSigma, Burlington, MA, USA), and immunoblotted with antibodies anti-HuR (Santa Cruz Biotechnology, Dallas, TX, USA), TRIM21, Chk2, Ubiquitin, Myc (Cell Signaling Technology, Danvers, MA, USA), His (BioBharati Life Sciences, India), AKT and RCHY1 (Proteintech, Rosemont, IL, USA), phosphoAkt (R&D Systems, Minneapolis, MN, USA), phosphoSerine (Santa Cruz Biotechnology), non-immune IgG (BioBharati), HRP conjugated β-actin (Genscript, Piscataway, NJ, USA) and GAPDH (Santa Cruz Biotechnology) antibodies as primary antibodies. HRP conjugated anti-mouse and anti-rabbit (Cell Signaling Technologies) were used as secondary antibodies. Chemiluminescent signal was detected using FemtolucentPlus HRP substrate (Geno Biosciences, St. Louis, MO, USA).

Nag S., Rahaman S., Guha A, & Ray P.S. (2023). An AKT1-and TRIM21-mediated phosphodegron controls proteasomal degradation of HuR enabling cell survival under heat shock. iScience, 26(4), 106307.

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Western Blot Analysis of Protein Targets

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Radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime Biotechnology, Shanghai, China) was used to extract total protein. The protein concentration was measured by a BCA protein assay kit (Thermo Fischer Scientific, Waltham, MA, USA). Equivalent proteins were electrophoresed via 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred onto a polyvinylidene fluoride (PVDF) membrane. After membranes were blocked in blocking buffer, it was followed by incubation with primary antibody at 4 °C overnight targeting hnRNPA1 (1:1000; Cell Signaling Technology, Beverly, MA, USA), Trim21 (1:1000; Cell Signaling Technology), PKM2 (1:1000; Cell Signaling Technology), PKM1 (1:1000; Cell Signaling Technology), IGF2BP3 (1:5000, Abcam, Cambridge, MA, USA), and GAPDH (1:3000; Cell Signaling Technology). Secondary antibody HRP-labeled rabbit IgG (1:5000; Cell Signaling Technology) was probed for 1 h at room temperature. The bands were detected by enhanced chemiluminescence.

Xie R., Liu L., Lu X., He C., Yao H, & Li G. (2023). N6-methyladenosine modification of OIP5-AS1 promotes glycolysis, tumorigenesis, and metastasis of gastric cancer by inhibiting Trim21-mediated hnRNPA1 ubiquitination and degradation. Gastric Cancer, 27(1), 49-71.

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Western Blot Antibody Characterization

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The western blot primary antibodies to flag (Sigma SAB4301135, 1:5000), HA (Abcam 9110, 1:1,000), Myc (Cell Signaling Technology 2272, 1:1,000), Actin (Beyotime AA128, 1:5000), Acetylated-lysine (Cell Signaling Technology 9441, 1:1,000), SHMT2(Santa Cruz 25064, 1:1,000), SIRT3 (Cell Signaling Technology 2627, 1:1,000), TRIM21(Cell Signaling Technology 92043, 1:1,000), K63-Ub (Abcam 179434, 1:1,000), COXIV (Cell Signaling Technology 3E11, 1:2,000), ATG5 (Cell Signaling Technology D5F5U, 1:1,000), Tublin (Beyotime AT819-1, 1:5000) were commercially obtained. Antibody specific to SHMT2 K95Ac was prepared commercially from immunizing rabbits at Shanghai Youke Biotechnology (antigen peptide: CLNNK (Ac) YSEGY-NH2, 1:200). NAM (Sigma 72345), Cycloheximide (CHX)(Sigma 01810), MG132(Sigma M8699), NH4Cl (Sangon Biotech), 3-methyladenine (3-MA)(Selleck, CAS:5142-23-4), sodium formate (Sigma 71539) are commercially obtained.

Wei Z., Song J., Wang G., Cui X., Zheng J., Tang Y., Chen X., Li J., Cui L., Liu C.Y, & Yu W. (2018). Deacetylation of serine hydroxymethyl-transferase 2 by SIRT3 promotes colorectal carcinogenesis. Nature Communications, 9, 4468.

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Western Blot Analysis of Protein Expression

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Cells were lysed using RIPA protein extraction reagent (Beyotime, Beijing, China) supplemented with phenylmethanesulfonyl fluoride (PMSF) (Riche, CA, USA). Approximately 50 µg of protein extracts were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred onto nitrocellulose membranes (Sigma), and incubated with specific antibodies. An enhanced chemiluminescent (ECL) chromogenic substrate was used to visualize the bands. The blots were developed with a chemiluminescence system, and GAPDH was used as a control. All the rabbit monoclonal antibodies were purchased from Cell Signaling Technology, including PD-L1 (1:5000), Bim, Bcl-2, cleaved caspases-3, cleaved caspases-8 (1:5000), TGF-β, TRIM21, NF-ƘB, AKT Phospho-AKT (Ser473) (1:2500), AKT (1:2500), Phospho-STAT3 (Tyr705) (1:3000), STAT3 (1:3000), E-cadherin, N-cadherin, Snail, and Vimentin (1:1000). Experimental groups are representative of at least 3 independent experiments.

Zhang H., Zhou F., Wang Y., Xie H., Luo S., Meng L., Su B., Ye Y., Wu K., Xu Y, & Gong X. (2020). Eliminating Radiation Resistance of Non-Small Cell Lung Cancer by Dihydroartemisinin Through Abrogating Immunity Escaping and Promoting Radiation Sensitivity by Inhibiting PD-L1 Expression. Frontiers in Oncology, 10, 595466.

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