BioLegend Product Reviews
Product: Purified anti-HA.11 Epitope Tag
Catalog No.: 901501
Ganesh Pusapati, Dr Stanford University

HA (mouse antibody) for Western Blotting
Overall: Product Quality: Ease of Use:
Over-expressed HA-tagged protein detection.
Experimental Design

Application: Western Blotting
Cells used: HEK293T cells
Brief Protocol: HA-IQCE protein was over-expressed in HEK293T cells for 24 hr. Whole cell extracts were prepared in RIPA lysis buffer. Protein estimation was done by BCA method. 50 µg of protein was loaded on an 8% Tris-Glycine gel and SDS-PAGE was performed. Proteins were transferred from the gel to a nitrocellulose membrane by semi-dry transfer method for 1 hr at room temperature. Protein transfer was visualized by Ponceau staining. The nitrocellulose membrane was washed well in ultra-pure water, blocked in blocking buffer (5% milk powder in 1xTBS-T) for 30 min at room temp and incubated for 16 hr at 4 deg with HA antibody (1:2000 dilution in blocking buffer). The membrane was washed thrice with 1x TBS-T and incubated with secondary antibody for 1 hr at room temperature (1:5000 dilution of Donkey anti-mouse IgG HRP in blocking buffer). The membrane was washed thrice in 1x TBS-T and incubated with 1 ml of ECL solution for 1 min at room temperature. Film was exposed for 2 sec and developed to visualize bands.
Results Summary: This antibody detects a HA-tagged IQCE protein (expected size ~130 kDa) with no background bands. Works very well for Western Blotting.
Additional Notes:
Data Image

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Publication Status

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