Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker

• WB

Lab

Western blot - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (AB18102)

Lanes 1 - 2 : Merged signal (red and green). Green - ab18102 observed at 300 kDa. Red - loading control, ab176560, observed at 50 kDa.

ab18102 was shown to recognize NES in wild-type HAP1 cells as signal was lost at the expected MW in NES knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NES knockout samples were subjected to SDS-PAGE. ab18102 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (ab18102) at 1/5000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

NES knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 177 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (AB18102)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of migrating tumor cells labelling Nestin with ab18102 at 2μg/ml. Cytoplasm and membrane positive staining shown.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (AB18102)

Immunohistochemistry of Anti-Nestin on formalin fixed paraffin embedded prostate nerve tissue using ab18102

• Flow Cyt

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Flow Cytometry - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (AB18102)

Overlay histogram showing SH-SY5Y cells stained with ab18102 (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 (ab18102, 1/200 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (AB18102)

Immunofluorescence staining of U251 cells (glioblastoma cell line) stained using ab18102(green) and bisbenzimide (blue)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (AB18102)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of xenograft tumor cells labelling Nestin with ab18102.

• WB

Supplier Data

Western blot - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (AB18102)

All lanes:

Western blot - Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker (ab18102) at 1/1000 dilution

Lane 1:

Total rat brain protein at 5 µg

Lane 2:

Human CNS progenitor cell protein at 1 µg

Lane 3:

Human CNS progenitor cell protein at 5 µg

Predicted band size: 177 kDa

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