Western blot analysis of the TAP tag was performed by loading the indicated amounts of GST-TAP-tag E. coli lysate and 10ul of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a low fluorescence PVDF membrane (Product # 22860) and blocked with SEA BLOCK Blocking Buffer (Product # 37527) for at least 1 hour. The membrane was probed with a DyLight 680-conjugated TAP tag monoclonal antibody (Product # MA1-108-D680) at a dilution of 1:1000 overnight at 4°C on a rocking platform, and washed in TBS-0.1% Tween-20. Fluorescent detection was performed using an IR imaging system.
|Tested species reactivity||Tag|
|Host / Isotype||Mouse / IgG1|
|Immunogen||KLH conjugated peptide representing the C-terminus of the TAP construct after TEV cleavage|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||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.
MA1108-D680 detects the Tev cleavage peptide sequence. MA1108-D680 has been successfully used for fluorescent detection of TAP in Western Blot.
DyLight 680 has an excitation/emission of 692/712nm.
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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