Characteristics
Tag: GST
Purity: > 95% by RP-HPLC and SDS-PAGE
Molecular Weight: 55,647 kDa
Physical State: Liquid
Quantity: 200 µg
Solubility: >30 mg/mL
Concentration: Variable (5-10 mg/mL )
Storage: -80°C, avoid freeze/thaw cycles
Application
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.
Benefits
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 trap-B 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).
Publications
(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.
14;14:200.
(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.
