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Ubiquitin monomer

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Ubiquitin is a small, highly conserved protein found in all eukaryotic cells. It functions as a post-translational modification, where it can be covalently attached to target proteins, thereby regulating their cellular function, localization, and stability. Ubiquitin monomers can be used in research and development applications to study ubiquitination processes.

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Lab products found in correlation

Dithiothreitol (dtt), by Merck Group (7 mentions) Bovine serum albumin (bsa), by Merck Group (6 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Maldi tof ms materials, by Bruker (3 mentions) Maldi tof ms 1536 anchorchip, by Bruker (2 mentions) Josd1 monoclonal antibody, by Thermo Fisher Scientific (2 mentions) Complete protease inhibitors cocktail tablets, by Roche (2 mentions) Hbx 41 108, by Bio-Techne (2 mentions) Pimozide, by Bio-Techne (2 mentions) Degrasyn wp1130, by Bio-Techne (2 mentions) L434078, by Merck Group (2 mentions) Pr 619, by Merck Group (2 mentions) P22077, by Merck Group (2 mentions) Doxycycline, by Merck Group (1 mentions) Hygromycin, by Thermo Fisher Scientific (1 mentions) Blasticidin, by Thermo Fisher Scientific (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Lds sample buffer, by Thermo Fisher Scientific (1 mentions) Instantblue coomassie stain, by Abcam (1 mentions) Glutathione sepharose, by GE Healthcare (1 mentions)

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Monomers Ubiquitin

7 protocols using ubiquitin monomer

1

Ubiquitin Ligation Assay Protocol

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Ubiquitin monomer, bovine serum albumin (BSA), Tris, dimethyl sulfoxide (DMSO), and DTT were purchased from Sigma-Aldrich. Ub-Lys-TAMRA-Gly was purchased from Boston Biochem (Boston, MA) (no. U-558). MALDI-TOF MS 1536 AnchorChip was purchased from Bruker Daltonics (Bremen, Germany).
The 2′,6′-dihydroxyacetophenone (DHAP) matrix was purchased from Tokyo Chemical Industry (product no. D1716). JOSD1 monoclonal antibody was purchased from Thermo Fisher (MA5-25365).
De Cesare V., Carbajo Lopez D., Mabbitt P.D., Fletcher A.J., Soetens M., Antico O., Wood N.T, & Virdee S. (2021). Deubiquitinating enzyme amino acid profiling reveals a class of ubiquitin esterases. Proceedings of the National Academy of Sciences of the United States of America, 118(4), e2006947118.
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2

Ubiquitin Topoisomer Preparation and Analysis

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Ubiquitin monomer, BSA, Tris and DTT were purchased from Sigma Aldrich. Di-Ubiquitin topoisomers (linear, K6, K11, K27, K29, K33, K48 and K63-linked ubiquitin-ubiquitin) were purchased from Boston Biochem. Complete protease inhibitors cocktail tablets were obtained from Roche. Protein G–Sepharose, glutathione–Sepharose and ECL reagents were from GE Healthcare. Doxycycline, DMSO, BSA, 3-amino, 1,2,4 triazole and benzamidine were from Sigma–Aldrich. PMSF was from Melford. Novex 4–12% polyacrylamide Bis-Tris gels, LDS sample buffer, puromycin, hygromycin, blasticidin and other tissue culture reagents were from Invitrogen Life Technologies. Instant Blue Coomassie stain was from Expedeon. PEI (polyethylenimine) was from Polysciences.
Conte C., Griffis E.R., Hickson I, & Perez-Oliva A.B. (2018). USP45 and Spindly are part of the same complex implicated in cell migration. Scientific Reports, 8, 14375.
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3

Ubiquitin Modification and Analysis

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Ubiquitin monomer, BSA, Tris, DTT and Gliotoxin were purchased from Sigma-Aldrich. MALDI TOF MS materials (targets, matrix and protein calibration mixture) were purchased from Bruker Daltonics (Bremen, Germany). PYR41 and Nutlin3A compounds were kindly provided by Sara Buhrlage (Dana-Farber Cancer Institute) and Compound-1 was kindly provided by Satpal Virdee (MRC PPU, Dundee).
De Cesare V., Johnson C., Barlow V., Hastie J., Knebel A, & Trost M. (2018). The MALDI-TOF E2/E3 Ligase Assay as Universal Tool for Drug Discovery in the Ubiquitin Pathway. Cell Chemical Biology, 25(9), 1117-1127.e4.
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4

Diubiquitin Topoisomer Characterization

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Ubiquitin monomer, BSA, Tris and DTT were purchased from Sigma-Aldrich. Diubiquitin topoisomers (M1, K6, K11, K27, K29, K33, K48 and K63-linked) were purchased from Boston Biochem (Boston, MA), additional K27 diubiquitin was produced in-house57 (link)58 , whereas all MALDI-TOF MS materials (targets, matrix and protein calibration mixture) were purchased from Bruker Daltonics (Bremen, Germany). PR-619 and P22077 (Calbiochem/Merck, Darmstadt, Germany) as well as HBX 41,108, pimozide and degrasyn/WP1130 (Tocris Bioscience, Bristol, UK) and L434078 (Sigma-Aldrich, St Louis, MO, USA) were purchased commercially. Febuxostat, SJB3-019A, compound 16, NSC 697923 and BAY 11-7082 were synthesized (Supplementary Table 3).
Ritorto M.S., Ewan R., Perez-Oliva A.B., Knebel A., Buhrlage S.J., Wightman M., Kelly S.M., Wood N.T., Virdee S., Gray N.S., Morrice N.A., Alessi D.R, & Trost M. (2014). Screening of DUB activity and specificity by MALDI-TOF mass spectrometry. Nature Communications, 5, 4763.
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5

Ubiquitin Topoisomers Biochemical Characterization

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Ubiquitin monomer, BSA, Tris, Triton X-100 and DTT, trimethyl-psoralen (TMP), hydroxyurea, angelicin and BrdU were purchased from Sigma-Aldrich. Di-ubiquitin topoisomers (linear, K6-, K11-, K27-, K29-, K33-, K48- and K63-linked ubiquitin–ubiquitin) were purchased from Boston Biochem. Complete protease inhibitors cocktail tablets were obtained from Roche. Tween-20, Colloidal blue staining kit and precast SDS–polyacrylamide Bis-Tris gels (Invitrogen). MMC was from Duchefa.
All recombinant proteins, plasmids and antibodies generated for the present study are available upon request and described in further detail on our reagents website (https://mrcppureagents.dundee.ac.uk/).
Perez-Oliva A.B., Lachaud C., Szyniarowski P., Muñoz I., Macartney T., Hickson I., Rouse J, & Alessi D.R. (2014). USP45 deubiquitylase controls ERCC1–XPF endonuclease-mediated DNA damage responses. The EMBO Journal, 34(3), 326-343.
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6

Quantitative Analysis of Diubiquitin Linkages

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Ubiquitin monomer, Bovine Serum Albumin (BSA), Tris and DTT were purchased from Sigma Aldrich. Diubiquitin topoisomers (M1, K6, K11, K27, K29, K33, K48 and K63-linked) were purchased from Boston Biochem (Boston, MA), additional K27 diubiquitin was produced in-house57 (link), 58 (link), whereas all MALDI-TOF MS materials (targets, matrix and protein calibration mixture) were purchased from Bruker Daltonics (Bremen, Germany). PR-619 and P22077 (Calbiochem/Merck, Darmstadt, Germany) as well as HBX 41,108, Pimozide and Degrasyn/WP1130 (Tocris Bioscience, Bristol, UK) and L434078 (Sigma-Aldrich, St. Louis, MO, USA) were purchased commercially. Febuxostat, SJB3-019A, Compound 16, NSC 697923 and BAY 11-7082 were synthesized (Supplementary Table 3).
Ritorto M.S., Ewan R., Perez-Oliva A.B., Knebel A., Buhrlage S.J., Wightman M., Kelly S.M., Wood N.T., Virdee S., Gray N.S., Morrice N.A., Alessi D.R, & Trost M. (2014). Screening of DUB activity and specificity by MALDI-TOF mass spectrometry. Nature Communications, 5, 4763.
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7

Ubiquitin Modification Characterization

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Ubiquitin monomer, BSA, Tris, DMSO and DTT were purchased from Sigma-Aldrich.
Ub-Lys-TAMRA was purchased from Boston Biochem (Boston, MA). MALDI-TOF MS 1536 AnchorChip was purchased from Bruker Daltonics (Bremen, Germany).
2',6'-dihydroxyacetophenone (2,6-DHAP matrix) was purchased from Tokyo Chemical Industry (Product Number：D1716). JOSD1 monoclonal antibody was purchased from ThermoFisher (MA5-25365).
Cesare V.D., Carbajo D., Mabbitt P.D., Antico O., Wood N.T., & Virdee S. (2020). Deubiquitinating Enzyme Amino Acid Profiling Reveals a Class of Ubiquitin Esterases.
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