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Western blot analysis of GST was performed by loading 25 ug of 293t cells transfected with GST-MART1 (lane 1), HepG2 cells (lane 2) and 293t cells transfected with 12Tag (lane 3) onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a GST Epitope Tag polyclonal antibody (Product # PA1-982A) at a dilution of 1:5000 (GST and 12Tag) and 1:1000 (HepG2) overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). The image shows the target bands on GST-MART1 (39kDa), HepG2 (26kDa) and 12Tag (45kDa), which are consistent with predicted molecular weights for each target. Exposure times were 2 min (lane 1), 1 min (lane 2) and 30 sec (lane 3).
|Tested species reactivity||Tag|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||GST-tagged fusion proteins.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000-1:5000|
* 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 1 publications below|
PA1-982A detects glutathione-S-transferase (GST).
PA1-982A has been successfully used in Western blot procedures to detect the presence of GST and GST-tagged proteins.
PA1-982A antigen is GST-tagged fusion protein.
Epitope tagging is a powerful and versatile strategy for detecting and purifying proteins expressed by cloned genes. To utilize this feature, protein expression vectors are typically engineered with a nucleotide sequence that encodes the peptide epitope tag. The gene of interest is cloned in-frame relative to the tag and, upon expression, the protein of interest is synthesized as a fusion protein with the peptide tag. Fusion protein detection and/or purification is mediated by highly specific antibodies to the engineered peptide, thus eliminating the need for antibodies to proteins from each newly cloned gene. Commonly used epitope tags include glutathione-S-transferase (GST), c-myc, 6-histidine (6X-His), FLAG®, green fluorescent protein (GFP), maltose binding protein (MBP), influenza A virus haemagglutinin (HA), b-galactosidase, and GAL4.
FLAG® and anti-FLAG® are registered trademarks of Sigma-Aldrich Biotechnology Co.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Identification of a novel phosphorylation site in protein phosphatase inhibitor-1 as a negative regulator of cardiac function.
PA1-982A was used in western blot to study the mechanism for cardiac contractility regulation
|Rodriguez P,Mitton B,Waggoner JR,Kranias EG||The Journal of biological chemistry (281:38599)||2006|