Immunofluorescent analysis of Chlamydia trachomatis FLAG-tagged IncA (blue) in HeLa cells. Cells were infected with a GFP-expressing strain of Chlamydia trachomatis containing FLAG-IncA under the control of the Tetracycline (Tet) promoter, and either left untreated (0ng/ml Tet) or treated with anhydrotetracycline (10ng/ml Tet). Cells were fixed with 2% paraformaldehyde in PBS for 30 minutes at room temperature, permeabilized and blocked in 10% goat serum, 0.1% BSA, 0.1% saponin in PBS pH 7.4 for 30 minutes at room temperature, and then probed with a FLAG Epitope Tag (DYKDDDDK) monoclonal antibody (Product # MA1-91878) at a dilution of 1:500 for 1 hour at room temperature. Cells were washed with PBS, and incubated with an AlexaFluor 647-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:2000 for 1 hour at room temperature. Images were taken on a fluorescent microscope using a 60X oil-immersion objective. Scale bars represent 20um. Data courtesy of the Innovators Program.
|Tested species reactivity||Tag|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Synthetic peptide (DYKDDDDK) coupled to KLH|
|Storage buffer||PBS, pH 7.2|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||3 ug|
|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 detects Flag tag from various samples.
similar epitope as the FLAG tag from Sigma
MA1-91878 has been successfully used in immunoprecipitation, Western blot and immunofluorescence applications.
The MA1-91878 immunogen is synthetic peptide (DYKDDDDK) coupled to KLH.
MA1-91878 may be stored at 4°C short term. Add 0.05% sodium azide if desired. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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.