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 an HRP-conjugated FLAG Epitope Tag monoclonal antibody (Product # MA1-91878-HRP) at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform and washed in TBS-0.1% Tween-20. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
|Tested species reactivity||Tag|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Synthetic peptide (DYKDDDDK) coupled to KLH.|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500 - 1:2000|
* 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|
similar epitope as the FLAG tag from Sigma
MA1-91878 detects Flag tag from various samples.
MA1-91878 has been successfully used in Western blot applications.
The MA1-91878 immunogen is synthetic peptide (DYKDDDDK) coupled to KLH.
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.
|Not Applicable||Not Cited||
Regulation of the human prostacyclin receptor gene by the cholesterol-responsive SREBP1.
MA1-91878-HRP was used in western blot to investigate the influence of cholesterol on human IP [(h)IP] expression
|Turner EC,Kinsella BT||Journal of lipid research (53:2390)||2012|