Characteristics
- Tag: GST
- Purity: > 95% by RP-HPLC and SDS-PAGE
- Molecular Weight: 26,98 kDa
- Physical State: Liquid
- Quantity: 200 µg
- Solubility: >40 mg/mL
- Concentration: Variable (5-20 mg/ml)
- Storage: -80°C, avoid freeze/thaw cycles
Applications
- Control in 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.
- Quantitative or semiquantitative methods using different supports.
Benefits
- Control background levels obtained in different affinity procedures and detection/quantification methods (6)
Examples of use
This GST has been used as control in different protocols to extract ubiquitylated proteins and in MS procedures. A step by step protocol using GST as control has been also reported (5).
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) 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.
(3) 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.
(4) 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.
(5) 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.
(6) 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.
