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 1.25 µg/ml of anti-human Nestin (clone 10C2) Brilliant Violet 421™ (magenta) overnight, followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes at 4°C. Nuclei were counterstained with DRAQ5™ (blue). The image was captured with a 20X 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 1.25 µg/ml of anti-human Nestin (clone 10C2) Brilliant Violet 421™ (magenta) overnight, followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes at 4°C. Nuclei were counterstained with DRAQ5™ (blue). The image was captured with a 20X objective.
Human paraffin-embedded ovary tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris-Buffered Saline 1X (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% Tween20 in PBS and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/ml of Alexa Fluor® 594 anti-human Cadherin 11 (clone 16G5) antibody (red) and Brilliant Violet 421™ anti-Nestin (clone 10C2) antibody (blue) overnight at 4°C. The image was scanned with a 10X objective and stitched with MetaMorph® software.
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 0.1 - 10 μg/ml is recommended. For immunohistochemical staining on formalin-fixed paraffin-embedded tissue sections, the suggested use of this reagent is 5.0 - 10 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.
Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.
This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.
Excitation Laser
Violet Laser (405 nm)
Application References
(PubMed link indicates BioLegend citation)
Jozef S, et al. 2012. Biomed. Pap. Med. Fac. 156:135. (IHC)
Product Citations
Rocktäschel P, et al. 2019. Epilepsy Behav. 101:106581. PubMed
Tamai K, et al. 2022. Cell Rep Methods. 2:100314. PubMed
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