Immuno-precipitation of GST-Shp2 SH2 domain (Product #87709) (Lane 2, 39kDa) was performed by mixing 2µg of purified fusion protein with1µg of mouse monoclonal antibody recognizing GST (Product # MA4-004) overnight on a rocking platform at 4°C. The immune-complex was then captured on 50µl Protein A/G Plus Agarose (Product #20423). Captured immune-complexes were then washed extensively and proteins eluted with 5X Reducing Sample Loading Dye (Product # 39000). Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel. IP efficiency was compared to a non-IP loading control consisting of 1ug GST-Shp2 SH2 domain (Lane1). Proteins were transferred to PVDF membrane and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing GST (Product # MA4-004) at a dilution of 1:1000 overnight rotating at 4°C. Membranes were then washed in TBS/0.1% Tween-20 and probed with Clean-blot IP detection reagent (Product #21230) at a dilution of 1:2000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Pierce Super Signal West Dura (Product #34075).
|Tested species reactivity||Tag|
|Host / Isotype||Mouse / IgG|
|Immunogen||Recombinant purified full-length GST|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:500 - 1:1000|
* 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.
MA4-004-BTIN has been successfully for Western blot and IP.
Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. The GST (Glutathione-S-Transferase) is a 26kDa protein encoded by the parasite Schistosoma japonicum, easily purified by affinity chromatography on immobilized glutathione. GST is utilized as a fusion protein with foreign proteins in a range of prokaryotic expression vectors, including the pGEX family of vectors. Antibodies against GST will detect the expressed GST protein or a GST-fusion protein, and can be utilized for immuno-purification of GST-fusion proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.