ICC staining of FITC anti-6-His Epitope Tag antibody (clone 6-His) on HeLa cells transfected with Posi-Tag plasmid. The cells were fixed with 4% PFA, and permeabilized with a buffer containing 0.5% Triton X-100, and blocked with 5% fetal bovine serum in PBS. The cells were then incubated with 10 µg/mL of the primary antibody overnight at 4°C. Nuclei were counterstained with DAPI, and the slide was mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 40X objective.
ICC staining of FITC anti-6-His Epitope Tag antibody (clone 6-His) on HeLa cells transfected with Posi-Tag plasmid. The cells were fixed with 4% PFA, and permeabilized with a buffer containing 0.5% Triton X-100, and blocked with 5% fetal bovine serum in PBS. The cells were then incubated with 10 µg/mL of the primary antibody overnight at 4°C. Nuclei were counterstained with DAPI, and the slide was mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 40X objective.
His tags are commonly used as a method of tagging proteins for detection or purification. This antibody is raised again a polyhistidine tag (6-His or hexa His-tag).
Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 5 - 10 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
Excitation Laser
Blue Laser (488 nm)
Application Notes
The sequence of the protein flanking the poly His tag does not influence antibody binding. This antibody was subsequently purified via a protein-G column.
Application References
(PubMed link indicates BioLegend citation)
Deinert K, et al. 2001. J. Biol. Chem. 276:6000. (WB)
Kojima H, et al. 2001. J. Biol. Chem. 276:19238. (IF) PubMed
Lapinski PE, et al. 2000. J. Biol. Chem. 275:6831. (WB)
Bartsch D, et al. 1998. Cell. 95:211. (IF)
Product Citations
Kojima H, et al. 2001. J Biol Chem. 276:19238-19243. PubMed
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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