Cell Signaling Technology

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DYKDDDDK Tag (9A3) Mouse mAb (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #8146

Applications Reactivity Sensitivity Isotype
W IP IHC-P IF-IC F All Transfected Only Mouse IgG1

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  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 (9A3) Mouse mAb detects exogenously expressed DYKDDDDK proteins in cells. The antibody recognizes the DYKDDDDK peptide, which is the same epitope recognized by Sigma's Anti-FLAG® antibodies.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic DYKDDDDK peptide.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, untransfected or transfected with either DYKDDDDK-tagged FoxG1, NPAS4, or Tyrosine Hydroxylase (TH), using DYKDDDDK Tag (9A3) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control (left) or ROS-DYKDDDDK Tag transfected (right), using DYKDDDDK Tag (9A3) Mouse mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Cos cells, untreated (blue) or transfected with a DYKDDDDK-tagged IRF3 contruct (green) using DYKDDDDK Tag (9A3) Mouse mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells transfected with a DYKDDDDK-tagged IRF3 construct using DYKDDDDK Tag (9A3) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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, 730-5.

Application References

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Companion Products

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

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