Western blot analysis of FLAG Epitope Tag was performed by loading various amounts of E. coli lysate containing a multi-epitope tagged protein 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 biotinylated FLAG Epitope Tag monoclonal antibody (Product # MA1-91878-BTIN) at a dilution of 1:1000 for 1 hour at room temperature, followed by Streptavidin-HRP (Product # 21126) at a dilution of 1:20,000 for 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).
|Tested species reactivity||Tag|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Synthetic peptide (DYKDDDDK) coupled to KLH.|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500 - 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.
MA1-91878-BTIN has been successfully used for Western blot, IF and IP applications.
similar epitope as the FLAG tag from Sigma
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. Tagging with xxxDDDDK may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, and in internal positions of the target protein. The small size of the epitope tag and its high hydrophilicity tend to decrease the possibility of interference with protein expression, proteolytic maturation, antigenicity and function. The enterokinase cleavage site allows it to be completely removed from the purified fusion proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.