Western blot analysis of TEV cleavage site was performed by loading the indicated amounts of TEV control protein and 5ul of Lane Marker Reducing Sample Buffer (Product # 39000) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a TEV cleavage site polyclonal antibody (Product # PA1-119) at a dilution of 1:500 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-rabbit IgG-HRP secondary antibody (Product # 32460) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
|Tested species reactivity||Tag|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide representing the cleavage site recognized by TEV enzyme|
|Purification||Antigen affinity chromatography|
|Storage buffer||BBS, pH 8.4|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||0.5 µg/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.
PA1-119 has been successfully used in Western blot applications.
The PA1-119 immunogen is a peptide representing the cleavage site recognized by TEV enzyme.
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. Tandem affinity purification (TAP) is an affinity purification method for isolation of TAP-tagged proteins together with associated proteins. The protocol involves the fusion of the "TAP tag" (typically a calmodulin binding peptide (CBP), a tobacco etch virus protease (TEV protease) cleavage site and Protein A) to the protein of interest. The TAP technique is useful in analyzing in vivo interactions.
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