Anti-Podoplanin antibody [EPR27193-535]

• Flow Cyt

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Flow Cytometry - Anti-Podoplanin antibody [EPR27193-535] (AB319138)

Flow cytometric analysis of NIH/3T3 (mouse embryonic fibroblast) (Right) / RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (Left) cells labelling Podoplanin with ab319138 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Low expression : RAW 264.7

• Flow Cyt

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Flow Cytometry - Anti-Podoplanin antibody [EPR27193-535] (AB319138)

Flow cytometric analysis of C2C12 (mouse myoblast) (Right) / Hepa1-6 (mouse hepatoma epithelial cell) (Left) cells labelling Podoplanin with ab319138 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Low expression : Hepa1-6

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Podoplanin antibody [EPR27193-535] (AB319138)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblast) cells labelling Podoplanin with ab319138 at 1/50 (10.12 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic with membranous staining in C2C12 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : Hepa1-6
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

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Western blot - Anti-Podoplanin antibody [EPR27193-535] (AB319138)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : Hepa1-6, RAW 264.7

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Podoplanin antibody [EPR27193-535] (ab319138) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

C2C12 (mouse myoblast) whole cell lysate at 20 µg

Lane 3:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 5:

Mouse lung tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 38 kDa,36 kDa

false

Exposure time: 180s