Cell Signaling Technology

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HA-Tag (6E2) Mouse mAb #2367

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

Applications Key:  W=Western Blotting  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

HA-Tag (6E2) Mouse mAb detects recombinant proteins containing the HA epitope tag. The antibody recognizes the HA-tag fused to either the amino or carboxy terminus of targeted proteins in transfected cells.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide containing the influenza hemagglutinin epitope (YPYDVPDYA) .

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untransfected (lane 1) or expressing HA-tagged Akt3 (lane 2) or HA-tagged Estrogen Receptor (ER) (lane 3), using HA-Tag (6E2) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded COS cells, untransfected (left), HA-Akt3 transfected (middle) or HA-Estrogen Receptor transfected (right), using an Akt antibody (top), an Estrogen Receptor antibody (middle) or HA-Tag (6E2) Mouse mAb (bottom).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS cells, untransfected (blue) or transfected with HA-Akt3 (green), using HA-Tag (6E2) Mouse mAb compared to a nonspecific negative control antibody (red).


IF-IC

IF-IC

Immunocytochemical staining of COS cells without transfection (left), expressing HA-tagged Akt3 (middle) or expressing HA-tagged Estrogen Receptor (right), using HA-Tag (6E2) Mouse mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS cells, mock-transfected (left) or transfected with HA-tagged protein (right), using HA-Tag (6E2) Mouse mAb (green). 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 HA tag is derived from an epitope of the influenza hemagglutinin protein, which has been used extensively as a general epitope tag in expression vectors (1).

  1. Field, J. et al. (1988) Mol. Cell. Biol. 8, 2159-2165.

Application References

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

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