Thrombin (MW 37kDa) is a Na⁺-activated, allosteric serine protease that plays opposing functional roles in blood coagulation. Thrombin recognition sequence and can be used to digest GST-tagged proteins.

p62 is cleaved from the agarose beads as well as the GST tag by addition of Thrombin (1 Unit/100 µg protein) at room temperature for 20 h.
Thrombin solution: Dissolve in PBS prechilled to 4°C. Swirl gently. Store solution in small aliquots at -80°C to preserve activity.
Cleavage buffer: PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 1.8 mM KH₂PO₄) pH 7.3.
Elution buffer: 50 mM Tris-HCl, 10 mM reduced glutathione, pH 8.0

西域 has not independently confirmed the accuracy of these methods. They are for reference only.

• . Golderman V, et al. Thrombin and the Protease-Activated Receptor-1 in Organophosphate-Induced Status Epilepticus. J Mol Neurosci. 2018 Dec 4.  [Content Brief]

  . Krenzlin H, et al. The Importance of Thrombin in Cerebral Injury and Disease. Int J Mol Sci. 2016 Jan 11;17(1).  [Content Brief]

  . Geetha T, et al. p62 serves as a shuttling factor for TrkA interaction with the proteasome. Biochem Biophys Res Commun. 2008 Sep 12;374(1):33-7.  [Content Brief]