HUVEC cells were fixed with 1% paraformaldehyde (PFA), permeabilized with 0.5% Triton X-100, and blocked with 5% fetal bovine serum for 30 minutes at room temperature. Then the cells were stained with 5 µg/ml of anti-human Nestin (clone 10C2) Alexa Fluor® 594 overnight, followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes at 4°C. Nuclei were counterstained with DAPI (blue). The image was captured with 40X objective.
HUVEC cells were fixed with 1% paraformaldehyde (PFA), permeabilized with 0.5% Triton X-100, and blocked with 5% fetal bovine serum for 30 minutes at room temperature. Then the cells were stained with 5 µg/ml of anti-human Nestin (clone 10C2) Alexa Fluor® 594 overnight, followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes at 4°C. Nuclei were counterstained with DAPI (blue). The image was captured with 40X objective.
Human paraffin-embedded kidney tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris-Buffered Saline 20X (1.0M, pH7.4) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween20 twice for five minutes , permeabilized with 0.5% Trito-X 100 for ten minutes and blocked with 5% FBS and 0.2% Gelatin for 30 minutes. Then, the tissue was stained with 5 µg/ml of purified anti-human CD133 (clone7) at 4°C overnight. On the next day, tissue was washed with PBS twice
Nestin is a class VI intermediate filament protein originally found in neuronal stem cells, and is required for survival and renewal of neural progenitor cells. In mixtures, Nestin forms heterodimers with vimentin or internexin, participating in the remodeling of the cell. Nestin was found to be expressed on vascular endothelial progenitor cells and various neoplasms. It is proposed to be a novel angiogenesis marker and cancer therapeutic target.
Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 1.0 - 5.0 µg/ml is recommended. For immunohistochemistry, a concentration range of 5.0 - 10 µg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
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