- Tag: GST
- Purity: > 95% by RP-HPLC and SDS-PAGE
- Molecular Weight: 56,476 kDa
- Physical State: Liquid
- Quantity: 200 µg
- Solubility: >30 mg/mL
- Concentration: Variable (5-10 mg/mL)
- Storage: -80°C, avoid freeze/thaw cycles
- Pull down of poly-ubiquitylated proteins from cell and tissue lysates using glutathione beads. Captured proteins can be analyzed by Western blot or Mass spectrometry with the appropriate adaptations.
- Protection of poly-ubiquitylated proteins from both deubiquitylation and degradation by the proteasome.
- Quantitative or semiquantitative methods using different supports.
- Up to 1000-fold higher affinity than the single UBD
- Avoid overexpression of tagged ubiquitin for pull downs
- Protect poly-ubiquitin chains without inhibitors specific to the proteasome or DUBs activity
- Replace ubiquitin specific antibodies for enrichment of poly-ubiquitylated proteins
Examples of use
UBI traps can be used to detect ubiquitylated cellular factors implicated in the regulation of signaling pathways regulating inflammation, oncogenesis, neurodegenerative diseases and multiple infections (4-8). The ubiquitylated status of these factors can be followed in response to a treatment and determine if cells are responding well (9-10). The ubiquitylation of crucial cellular factors can also be followed after infection with various microorganisms (11). Among other examples. A UBI traps step by step protocol has also been reported (12).
(1) Efficient protection and isolation of ubiquitylated proteins using tandem ubiquitin-binding entities. Hjerpe R, Aillet F, Lopitz-Otsoa F, Lang V, England P, Rodriguez MS.EMBO Rep. 2009 Nov;10(11):1250-8.
(2) Polyubiquitin-sensor proteins reveal localization and linkage-type dependence of cellular ubiquitin signaling. Sims JJ, Scavone F, Cooper EM, Kane LA, Youle RJ, Boeke JD, Cohen RE.Nat Methods. 2012 Feb 5;9(3):303-9.
(3) Using Ubiquitin Binders to Decipher the Ubiquitin Code. Mattern M, Sutherland J, Kadimisetty K, Barrio R, Rodriguez MS.Trends Biochem Sci. 2019 Jul;44(7):599-615.
(4)Role of monoubiquitylation on the control of IκBα degradation and NF-κB activity. Da Silva-Ferrada E, Torres-Ramos M, Aillet F, Campagna M, Matute C, Rivas C, Rodríguez MS, Lang V. PLoS One. 2011;6(10):e25397.
(5) Tetramerization-defects of p53 result in aberrant ubiquitylation and transcriptional activity. Lang V, Pallara C, Zabala A, Lobato-Gil S, Lopitz-Otsoa F, Farrás R, Hjerpe R, Torres-Ramos M, Zabaleta L, Blattner C, Hay RT, Barrio R, Carracedo A, Fernandez-Recio J, Rodríguez MS, Aillet F. Mol Oncol. 2014 Jul;8(5):1026-42.
(6) Inhibition of the proteasome and proteaphagy enhances apoptosis in FLT3-ITD-driven acute myeloid leukemia. Lopez-Reyes RG, Quinet G, Gonzalez-Santamarta M, Larrue C, Sarry JE, Rodriguez MS. FEBS Open Bio. 2021 Jan;11(1):48-60.
(7) Analysis of defective protein ubiquitylation associated to adriamycin resistant cells. Lang V, Aillet F, Xolalpa W, Serna S, Ceccato L, Lopez-Reyes RG, Lopez-Mato MP, Januchowski R, Reichardt NC, Rodriguez MS. Cell Cycle. 2017;16(24):2337-2344.
(8) Activation of AHR mediates the ubiquitination and proteasome degradation of c-Fos through the induction of Ubcm4 gene expression. Mejía-García A, González-Barbosa E, Martínez-Guzmán C, Torres-Ramos MA, Rodríguez MS, Guzmán-León S, Elizondo G. Toxicology. 2015 Nov 4;337:47-57.
(9) Activation of aryl hydrocarbon receptor regulates the LPS/IFNgamma-induced inflammatory response by inducing ubiquitin-proteosomal and lysosomal degradation of RelA/p65. Domínguez-Acosta O, Vega L, Estrada-Muñiz E, Rodríguez MS, Gonzalez FJ, Elizondo G. Biochem Pharmacol. 2018 Sep;155:141-149.
(10) Efficient monitoring of protein ubiquitylation levels using TUBEs-based microarrays. Serna S, Xolalpa W, Lang V, Aillet F, England P, Reichardt N, Rodriguez MS. FEBS Lett. 2016 Aug;590(16):2748-56.
(11) New insights into host-parasite ubiquitin proteome dynamics in P. falciparum infected red blood cells using a TUBEs-MS approach. Mata-Cantero L, Azkargorta M, Aillet F, Xolalpa W, LaFuente MJ, Elortza F, Carvalho AS, Martin-Plaza J, Matthiesen R, Rodriguez MS. J Proteomics. 2016 Apr 29;139:45-59.
(12) Isolation of ubiquitylated proteins using tandem ubiquitin-binding entities. Aillet F, Lopitz-Otsoa F, Hjerpe R, Torres-Ramos M, Lang V, Rodríguez MS.Methods Mol Biol. 2012;832:173-83.