GST Cleavage Capture Kit

The GST Cleavage Capture Kit allows easy release of free recombinant protein from their GST-tag when a Factor Xa recognition sequence is present.\n\nWhile GST-fusion proteins allow convenient affinity purification of many proteins of interest, the GST-tag may become inconvenient in various downstream applications. This GST Cleavage Capture Kit is designed for efficient removal of such GST-tags in fusion proteins where the amino acid sequence Ile-Glu(or Asp)-Gly-Arg-//-X is present in between the protein of interest and GST. This sequence is efficiently cleaved by the protease Factor Xa, and the two cleavage products (GST and protein of interest) can be easily separated. This cleavage reaction is usually completed within a few hours at temperatures between 4 degrees C to 37 degrees C, and the free GST-tag is removed by GSH-gel treatment.\n\nKit Components:\nFactor Xa stock solution:\n10ug in 10ul Tris Buffer solution (20mM, ph 7.4) activity: 20,000 IU/mg\nPlease note that dilutions to a lower concentration than 1ug/10ul may result in loss of protein due to adsorption.\n\nInhibitors:\nPhenylmethylsulfonyl fluoride (PMSF) 20ul 100x stock\nDithiothreitol (DTT) 20ul 100x stock\nChloromethylketone in NaCl\nBoc-DPhe-Arg(Z2)-CH2-Cl dissolve in 25ul Cleavage Buffer\nCL-Glutathione ChroMatrixTM 2x1ml\nWash Buffer (A) 50ml, contains 10mM mercaptoethanol\nWash Buffer (B) 10ml, contains 10mM mercaptoethanol\nElution Buffer (C) 25ml, contains 5mM DTT\nCleavage Buffer (D) 20ml, contains 1mM DTT\nElution Buffer (E) 5ml, contains 5mM GSH\n\nMaterials Required But Not Supplied:\nmicropipettes\nequipment for GSH affinity chromatography\ncentrifuge\nSDS-PAGE and staining solutions\n\nStorage & Stability:\nQuality guaranteed for 12 months Store at 4 degrees C, keep Factor Xa at -80 degrees C

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