Cell Signaling Technology

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DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate) #5407

Applications Reactivity Sensitivity Source
IF-IC F All Transfected Only Rabbit

Applications Key:  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

DYKDDDDK Tag Antibody (Alexa Fluor® 488 Conjugate) detects exogenously expressed DYKDDDDK proteins in cells. The antibody recognizes the DYKDDDDK peptide (the same epitope recognized by Sigma's Anti-FLAG® antibodies) fused to either the amino- or carboxy-terminus of targeted proteins. The binding specificity of this antibody is NOT dependent on the presence of divalent metal cations.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic DYKDDDDK peptide. Antibodies are purified by protein A and peptide affinity chromatography.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS cells, either untransfected (blue) or transfected with a DYKDDDDK-tagged fusion protein, using DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate) (green).

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS cells transiently transfected with a DYKDDDDK-tagged fusion protein using DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate) (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in cells transiently transfected with a DYKDDDDK-tagged fusion protein.

Background

Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein's biochemical properties.

The DYKDDDDK peptide has been used extensively as a general epitope tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion (1).

  1. Brizzard, B. L. et al. (1994) Biotechniques 16, 730-735.

Application References

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Companion Products

Anti-FLAG® is a registered trademark of Sigma-Aldrich.

For Research Use Only. Not For Use In Diagnostic Procedures.

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