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Schistosoma japonicum Glutathione S-transferase / GST Insect Cell Lysate (WB positive control)

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Schistosoma japonicum GST Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Baculovirus-Insect cells
Product Description:Baculovirus-Insect Cell lysate that Glutathione S-transferase / GST transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the Glutathione S-transferase (P08515) (Met 1-Lys 218), with additional 11 amino acids at the C-terminus, was expressed and purified.
Predicted N Terminal:Met 1
Molecule Mass:The recombinant Glutathione S-transferase (GST) consists of 229 amino acids and predicts a molecular mass of 26.9 kDa. The apparent molecular mass of GST is approximately 28 kDa in SDS-PAGE under reducing conditions.
Species:Schistosoma japonicum
Schistosoma japonicum GST Transfected / Overexpression Cell Lysate Usage Guide
Preparation Method:Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer:Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
Recommend Usage:1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
Glutathione S-transferase/GST Background

Genetic engineers have used glutathione S-transferase to create the GST gene fusion system. This system is used to purify and detect proteins of interest. In a GST gene fusion system, the GST sequence is incorporated into an expression vector alongside the gene sequence encoding the protein of interest. Induction of protein expression from the vector's promoter results in expression of a fusion protein: the protein of interest fused to the GST protein. This GST-fusion protein can then be purified from cells via its high affinity for glutathione. GST is commonly used to create fusion proteins. The tag has the size of 220 amino acids (roughly 26 KDa), which, compared to other tags like the myc- or the FLAG-tag, is quite big. However, many commercially-available sources of GST-tagged plasmids include a thrombin domain for cleavage of the GST tag during protein purification.

Schistosoma japonicum Glutathione S-transferase/GST References
  • Douglas KT . et al., 1987, Adv Enzymol Relat Areas Mol Biol.?59: 103- 67.?
  • Beckett GJ. et al., 1993, ?Adv Clin Chem. 30: 281-380.
  • Wilce MC. et al., 1994, ?Biochim Biophys Acta. 1205?(1): 1-18.
  • Leaver MJ. et al., 1998, ?Marine Environmental Research. 46?(1-5): 71-4.
Size / Price
Catalog: 11213-HNABL-300
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