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Western blot analysis of GST was performed by loading 30 µg of whole cell extracts of untransfected HEK-293 cell lysate (lane1) and HEK-293 transiently over expressing GST-GAD45 (lane2) using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. GST was detected at ~90 kDa using GST Mouse Monoclonal Antibody (136700) at 0.5-1 µg/ml in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse IgG - HRP Secondary Antibody (626520) at 1:4000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Tag|
|Published species reactivity||Rat , Eubacteria , Human|
|Host / Isotype||Mouse / IgG2b, kappa|
|Immunogen||GST protein from Schistosoma japonicum expressed from the pGEX vector.|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Immunofluorescence (IF)||Assay Dependent|
|Immunoprecipitation (IP)||2-5 ug|
|Western Blot (WB)||1.0 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 3 publications below|
GST (Glutathione S-transferase) is a 26kDa protein, present in eukaryotes and in prokaryotes, where they catalyze a variety of reactions. Glutathione Stransferase is used to create the GST gene fusion system. This GST-fusion protein can then be purified from cells via its high affinity for glutathione. Fusion Proteins offer an important biological assay for direct protein-to-protein interactions. The GST gene fusion system has been introduced into numerous expression vectors.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Raf kinase inhibitory protein regulates Raf-1 but not B-Raf kinase activation.
||Trakul N,Menard RE,Schade GR,Qian Z,Rosner MR||The Journal of biological chemistry (280:24931)||2005|
Two heme binding sites are involved in the regulated degradation of the bacterial iron response regulator (Irr) protein.
||Yang J,Ishimori K,O'Brian MR||The Journal of biological chemistry (280:7671)||2005|
The Erbin PDZ domain binds with high affinity and specificity to the carboxyl termini of delta-catenin and ARVCF.
||Laura RP,Witt AS,Held HA,Gerstner R,Deshayes K,Koehler MF,Kosik KS,Sidhu SS,Lasky LA||The Journal of biological chemistry (277:12906)||2002|
Glutathione-S-Transferase, GST protein, GST Tag