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Immunofluorescence (red) with Anti-His Tag Antibody on His-tag fusion protein in HEK293 cells. Left hand panel: untransfected control. Right hand panel: transfected.?Counterstained with DAPI (blue).

Standard ladder containing five different His-tagged proteins; untransfected (1), HEK293 cells transferred with His-tagged protein vector (2).

Comparison between Anti-His tag (HIS.H8 / EH158) mAb with 2 different vendor Abs, probed against a standard ladder containing five different His-tagged proteins. All Ab dilutions are 1:2000 (0.5μg/ml).

Western blot analysis of a 6x His-tagged protein was performed by loading 15μg of cell lysates from SF9 cells expressing an 11kD 6x-His tagged protein per well onto an SDS-PAGE gel. Proteins were transferred to a nitrocellulose membrane, and blocked in blocking buffer for 1 hour at room temperature. The membrane was probed with Anti-His Tag Antibody at a dilution of 1:500 overnight at 4 degrees celsius, washed in TBST, and probed with an Anti-Mouse IgG secondary antibody conjugated to a near-IR dye. Results were visualized using a near-IR imager.