Immunofluorescent analysis of HeLa cells transfected with a construct containing a FLAG Epitope Tag. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a DyLight 680-conjugated FLAG Epitope Tag monoclonal antibody (Product # MA1-91878-D680) at a dilution of 1:25 for at least 1 hour at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
|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 *|
|Immunofluorescence (IF)||1:25 - 1:100|
|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-D680 has been successfully used in immunofluorescence and Western blot applications. DyLight 680 has an excitation/emission of 692/712 nm.
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.