Total lysates (15 µg protein) from HeLa (Low expression negative control) and PC-3 (Positive control) were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:500 (1 µg/ml) purified anti-Nestin antibody, clone 10C2. Proteins were visualized using chemiluminescence detection by incubation with HRP Goat anti-Mouse secondary antibody (Cat. No. 405306, 1:3000 dilution). Direct-Blot™ HRP anti-β-actin antibody was used as a loading control (Cat. No. 643807, 1:8000 dilution).
Total lysates (15 µg protein) from HeLa (Low expression negative control) and PC-3 (Positive control) were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:500 (1 µg/ml) purified anti-Nestin antibody, clone 10C2. Proteins were visualized using chemiluminescence detection by incubation with HRP Goat anti-Mouse secondary antibody (Cat. No. 405306, 1:3000 dilution). Direct-Blot™ HRP anti-β-actin antibody was used as a loading control (Cat. No. 643807, 1:8000 dilution).
NTERA-2 cells were fixed with 2% paraformaldehyde (PFA) for 10 minutes, permeabilized with 0.5% Triton X-100 for 5 minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 2 µg/mL anti-Nestin Antibody (10C2) and followed by DyLight™ 594 goat anti-mouse IgG (red) for 1 hour at room temperature. Actin filaments were labeled with Alexa Fluor® 488 Phalloidin (green). Nuclei were counterstained with DAPI (blue). The image was captured with a 40X objective.
NTERA-2 cells were fixed with 2% paraformaldehyde (PFA) for 10 minutes, permeabilized with 0.5% Triton X-100 for 5 minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 2 µg/mL anti-Nestin Antibody (10C2) and followed by DyLight™ 594 goat anti-mouse IgG (red) for 1 hour at room temperature. Actin filaments were labeled with Alexa Fluor® 488 Phalloidin (green). Nuclei were counterstained with DAPI (blue). The image was captured with a 40X objective.
IHC staining of Purified anti-Nestin (clone 10C2) on formalin-fixed paraffin-embedded human kidney tissue. Following antigen retrieval with 1X Citrate Buffer (Cat. No. 420902), the tissue was incubated either without (panel A) or with (panel B) Purified anti-Nestin (clone 10C2), followed by incubation with Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 10X objective. Scale bar: 50 µm
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 Western blotting. For Western blotting, the suggested use of this reagent is 0.5 - 2.0 µg per mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1.0 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
Application References
Jozef S, et al. 2012. Biomed. Pap. Med. Fac. 156:135. (IHC)
Product Citations
Salgado B, et al. 2023. Microorganisms. 11:. PubMed
Di Stefano B, et al. 2019. Cell Stem Cell. 25:622. PubMed
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