Anti-Nestin Antibody (S1409)

Fluorescent confocal image of SH-SY5Y cells stained with Nestin (S1409) antibody. SH-SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0. 2%, 30 min). Cells were then incubated with A00806-1 Nestin (S1409) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). Note the highly specific localization of the Nestin (S1409) immunosignal to the intermediate filaments, supported by Human Protein Atlas Data (http://www. proteinatlas. org/ENSG00000132688).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0. 1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma cell line) cells labeling Nestin with A00806-1 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-rabbit IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoskeleton staining on U-2 OS cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin at 1/100 dilution (red). The nuclear counter stain is DAPI (blue).

All lanes : Anti-Nestin Antibody (S1409) at 1:2000 dilution
Lane 1: U-251 MG whole cell lysate
Lane 2: SH-SY5Y whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution.
Predicted band size : 177 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.

Anti-Nestin Antibody (S1409) at 1:2000 dilution + Hela whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution.
Predicted band size : 177 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.

A00806-1 staining Nestin in Human heart tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

A00806-1 staining Nestin in human brain tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

A00806-1 staining Nestin in human cerebellum tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.