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488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor?

594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPIwas used to stain the cell nuclei (blue) at a concentration of 1.43?M. Immunohistochemistry (Formalin/PFA-fixed paraffin- embedded sections) - Anti-Nestin antibody [SP103] (ab105389) Malchenko, S. et al PLoSOne.

2017 Mar 1;

12(3):e0173106. doi: 10.1371/journal.pone.0173106. eCollection

2017 BTIC markers in our CNS-PNET model tumors Formalin-fixed, paraffin-embedded mouse primitive neural ectodermal tumor tissue was stained for Nestin using ab105389 at 1/30 dilution in immunohistochemical analysis. Insert x20 magnification. TU-tumor;

P-parenchyma;

V-ventricle;

CP-choroid plexus (From Figure 3C of Malchenko et al) ? ?

3 Immunohistochemistry (Formalin/PFA-fixed paraffin- embedded sections) - Anti-Nestin antibody [SP103] (ab105389) Formalin-fixed, paraffin-embedded human endometrium tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis. Western blot - Anti-Nestin antibody [SP103] (ab105389) Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution + HeLa cell lysate Predicted band size:

177 kDa Flow Cytometry - Anti-Nestin antibody [SP103] (ab105389) Overlay histogram showing SH-SY5Ycells stained with ab105389 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for

20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab105389, 1/50 dilution) for

30 min at 22?C. The secondary antibody used was DyLight?

488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for

30 min at 22?C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1?g/1x10 cells) used under the same conditions. Acquisition of >

5,000 events was performed.

6 4 Immunohistochemistry (Formalin/PFA-fixed paraffin- embedded sections) - Anti-Nestin antibody [SP103] (ab105389) Formalin-fixed, paraffin-embedded human colon tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical anal........