Cell Signaling Technology

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DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368

Applications Reactivity Sensitivity Source
W IP IHC-P IF-IC F All Transfected Only Rabbit

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 Antibody 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.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS or 293T cells, mock transfected (lane 1) or expressing DYKDDDDK-tagged p73 (lane2), DYKDDDDK-tagged FKHRL1 (lane 3) or truncated DYKDDDDK-tagged huntingtin (lane 4), using DYKDDDDK Tag Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded COS cells, control (left) or DYKDDDDK-FKHRL1 transfected (right), using DYKDDDDK Tag Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of DYKDDDDK Tag Rabbit antibody in Cos cells transfected with a Myc-DDK-tagged SUMF1 construct and gated on transfected (green) or untransfected (blue) cells using Myc-Tag (9B11) Mouse mAb (Alexa Fluor® Conjugate) #2279.


IF-IC

IF-IC

Confocal immunofluorescent analysis of COS cells, untransfected (right) or transfected with a construct overexpressing DYKDDDDK-tagged FKHRL1 (left) using DYKDDDDK Tag Antibody.

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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