Hela cells transfected with multi-tag plasmid were fixed with cold methanol for 15 minutes at -20°C and then blocked with 5% FBS for 60 minutes. Cells were then stained with either (A) 1:500 (1 µg/ml) Alexa Fluor® 488 mouse IgG1, κ isotype control antibody (Cat no. 400133), or (B, C) 1:500 (1 µg/ml), (D) 1:1000 (0.5 µg/ml) or (E) 1:2500 (0.2 µg/ml) Alexa Fluor® 488 anti-His antibody (green), and incubated overnight at 4°C. Nuclei were counterstained with DAPI (blue, Cat no. 422801). The image was captured with a 60X objective using KEYENCE BZ-X700 fluorescence microscope. Exposure time (S): (A) 1/15, (B, D, E) 1/50, (C) 1/150.
Hela cells transfected with multi-tag plasmid were fixed with cold methanol for 15 minutes at -20°C and then blocked with 5% FBS for 60 minutes. Cells were then stained with either (A) 1:500 (1 µg/ml) Alexa Fluor® 488 mouse IgG1, κ isotype control antibody (Cat no. 400133), or (B, C) 1:500 (1 µg/ml), (D) 1:1000 (0.5 µg/ml) or (E) 1:2500 (0.2 µg/ml) Alexa Fluor® 488 anti-His antibody (green), and incubated overnight at 4°C. Nuclei were counterstained with DAPI (blue, Cat no. 422801). The image was captured with a 60X objective using KEYENCE BZ-X700 fluorescence microscope. Exposure time (S): (A) 1/15, (B, D, E) 1/50, (C) 1/150.
This antibody was raised against linear 8XHis tag. It reacts with His tag protein, either C, N, or internal His6 and up repeats. It can be used for Western blotting to detect linear His repeat (His6 or more).
Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 0.2 - 0.5 μg/ml (Dilution 1:1000-1:2500) is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.
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