HA Antibody for Western Blotting

Biochemistry
Stanford University
Senior Research Scientist

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Company:

BioLegend

Product Name:

Purified anti-HA.11 Epitope Tag Antibody

Catalog Number:

901501

This antibody was used to test the expression of a HA tagged protein overexpressed in HEK293T cells. The end goal of this experiment was to use this antibody to test the interaction of a HA tagged protein with a GFP tagged bait protein.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T cells

Primary Incubation

1:2000 dilution in blocking agent for 16 hr at 4 deg

Blocking Agent

5% milk powder in 1xTBS-T (0.1% Tween)

Secondary Incubation

Donkey anti-mouse IgG HRP used at 1:5000 dilution in blocking agent for 1 hr at room temp

Tertiary Incubation

None

Detection

ECL; exposure time 3 sec

Results Summary

HA-tagged Evc protein was over-expressed in HEK293T cells for 24 hr. Whole cell extracts were prepared in RIPA lysis buffer and 50 µg of the lysate for loaded on a 8% Tris-Glycine gel and SDS-PAGE was performed. The gel was transferred onto a nitrocellulose membrane and blotted with HA antibody. HA-Evc runs at ~130 kDa and the antibody nicely detects the protein.

Additional Notes

None

Related Categories

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Summary

The Good

Specific signal was observed

The Bad

None

The Bottom Line

Works wonderfully with no background issues

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