Cell Signaling Technology

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HA-Tag (6E2) Mouse mAb (Alexa Fluor® 647 Conjugate) #3444

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

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

HA-Tag (6E2) Mouse mAb (Alexa Fluor® 647 Conjugate) 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). The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS cells, untransfected (blue) or transfected with HA-tagged DLL1 (green), using HA-Tag (6E2) Mouse mAb (Alexa Fluor® 647 Conjugate).

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS cells, untransfected (left) or transfected with an HA-tagged protein (right), using HA-Tag (6E2) Mouse mAb (Alexa Fluor® 647 Conjugate) (blue pseudocolor) and Di-Methyl-Histone H3 (Lys27) Antibody #9755 (green). Actin filaments were labeled with DY-554 phalloidin (red).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in cells transfected with HA-tagged 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 HA tag is derived from an epitope of the influenza hemagglutinin protein, which has been extensively used as a general epitope tag in expression vectors (1).

  1. Field, J. et al. (1988) Mol Cell Biol 8, 2159-65.

Application References

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Companion Products

The Alexa Fluor® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc., for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents.Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

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

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