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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||Tag, Rat, Bacteria, Mouse, 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 4 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.
|Rat||Not Cited||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|
|Connexin43 associated with an N-cadherin-containing multiprotein complex is required for gap junction formation in NIH3T3 cells.||Wei CJ,Francis R,Xu X,Lo CW||The Journal of biological chemistry (280:19925)||2005|
|Bacteria||Not Cited||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|
|Human||Not Cited||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|