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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 PVDF membrane and blocked with StartingBlock T20 Blocking Buffer (Product # 37543) for at least 1 hour. The membrane was probed with a biotin-conjugated TAP tag monoclonal antibody (Product # MA1-108-BTIN) at a dilution of 1:1000 overnight at 4°C on a rocking platform, followed by Streptavidin-HRP (Product # 21126) at a dilution of 1:1000 for 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). Images were acquired on a Thermo Scientific myECL Imager (Product # 62236).
|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-BTIN detects the Tev cleavage peptide sequence. MA1108-BTIN has been successfully used for Streptavidin-HRP mediated detection of TAP in Western Blot.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.