HeLa cells transiently transfected with His tag fused protein were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with 1:1000 diluted (0.5 µg/ml) Alexa Fluor® 647 anti- His tag antibody (red). Nuclei were counterstained with DAPI (blue, Cat. No. 422801). The image was captured with a 40X objective. For negative control and titration data, refer to the supplemental data.
HeLa cells transiently transfected with His tag fused protein were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with 1:1000 diluted (0.5 µg/ml) Alexa Fluor® 647 anti- His tag antibody (red). Nuclei were counterstained with DAPI (blue, Cat. No. 422801). The image was captured with a 40X objective. For negative control and titration data, refer to the supplemental data.
HeLa cells transiently transfected with His tag fused protein or non-transfected were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with 1:250 (2 µg/ml), 1: 1000 (0.5 µg/ml) and 1:2500 diluted (0.2 µg/ml) Alexa Fluor® 647 anti- His Tag antibody (red). Nuclei were counterstained with DAPI (blue, Cat. No. 422801). The image was captured with a 40X objective using KEYENCE BZ-X700 fluorescence microscope.
This antibody was raised against a linear 8XHis tag. It reacts with His-tagged protein with C, N or internal 6XHis tags or longer. It can be used for western blotting to detect linear His repeats (6XHis or more). It can also be used for ICFC and ICC staining of His-tagged proteins.
Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 0.2 - 2.0 μg/ml (1:250 - 1:2500 dilution) is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
This antibody was raised against a linear 8X His tag. It reacts with proteins displaying repeats of 6 Histidines (His6) or more in either internal or C and N terminal regions. It is suitable for ICFC.
Clone J095G46 has been verified for immunocytochemistry (ICC).
Product Citations
Zhang B, et al. 2021. Nat Biomed Eng. 5:1288. PubMed
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