100 µg of protein G purified antibody (1 mg/ml) in PBS, 1 mg/ml BSA, and 0.05% sodium azide.
Western Blot (WB)
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Product Specific Information
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.
GST Antibody (MA4-004) in IP
Immunoprecipitation with anti-GST Monoclonal Antibody [8-326] (MA4-004)
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).
GST Antibody (MA4-004) in WB
Western Blot with anti-GST Monoclonal Antibody [8-326] (MA4-004)
Western blot analysis of GST-Abl SH2 domain (Lane 1, 35kDa) (Product # 87702) and GST-Grb2 (Lane 2, 38kDa) SH2 domain (Product #87700) was performed by loading 2ug of each purified fusion protein onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/0.1%TBST for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing GST clone 8-826 (Product # MA4-004) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were then washed in TBS/0.1%Tween-20 and probed with a goat anti-mouse-HRP secondary antibody (Product #31430) at a dilution of 1:25000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Pierce SuperSignal West Dura (Product #34075).
MA4-004 was used in western blot to investigate the association between WSP-like protein WSP-0284 and Brugia malayi proteins
Int J Parasitol. 2011 Aug 15;41(10):1053-61. "Interaction of a Wolbachia WSP-like protein with a nuclear-encoded protein of Brugia malayi."
Author(s): Melnikow E, Xu S, Liu J, Li L, Oksov Y, Ghedin E, Unnasch TR, Lustigman S
Number of Citations: 0
(See PubMed article )
(This product is for In Vitro experimental use only. Not for resale without express authorization.)