Anti-Syntenin antibody

• WB

Lab

Western blot - Anti-Syntenin antibody (AB19903)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : Syntenin knockout HAP1 whole cell lysate (20 μg)
Lane 3 : A549 whole cell lysate (20 μg)
Lane 4 : HeLa whole cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab19903 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab19903 was shown to recognize Syntenin in wild-type HAP1 cells as signal was lost at the expected MW in Syntenin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Syntenin knockout samples were subjected to SDS-PAGE. ab19903 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Syntenin antibody (ab19903)

Predicted band size: 32 kDa

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Syntenin antibody (AB19903)

ICC/IF image of ab19903 stained human HEK 293 cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab19903, 1μg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

• WB

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Western blot - Anti-Syntenin antibody (AB19903)

ab19903 detects a band of the expected size (32kDa) in HEK-293 cell lysate. This band is partially quenched by the addition of ab20431. No band is observed in human heart lysate.

All lanes:

Western blot - Anti-Syntenin antibody (ab19903) at 1 µg/mL

Lane 1:

HEK-293 cell lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

HEK-293 cell lysate at 20 µg with Mouse Syntenin peptide (<a href='/en-us/products/unavailable/mouse-syntenin-peptide-ab20431'>ab20431</a>)

Lane 4:

Human heart tissue lysate at 20 µg with Mouse Syntenin peptide (<a href='/en-us/products/unavailable/mouse-syntenin-peptide-ab20431'>ab20431</a>)

Predicted band size: 32 kDa

Observed band size: 32 kDa

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• WB

Unknown

Western blot - Anti-Syntenin antibody (AB19903)

ab19903 detects a band of the expected size (32kDa) in both mouse and rat heart lysate. This band is quenched completely by the addition of the immunizing peptide in the mouse lysate and is partially quenched by the addition of ab20431 in the rat heart lysate.

All lanes:

Western blot - Anti-Syntenin antibody (ab19903) at 1 µg/mL

Lane 1:

Mouse heart lysate at 20 µg

Lane 2:

Rat heart lysate at 20 µg

Lane 3:

Mouse heart lysate at 20 µg with Mouse Syntenin peptide (<a href='/en-us/products/unavailable/mouse-syntenin-peptide-ab20431'>ab20431</a>)

Lane 4:

Rat heart lysate at 20 µg with Mouse Syntenin peptide (<a href='/en-us/products/unavailable/mouse-syntenin-peptide-ab20431'>ab20431</a>)

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-ab6721'>ab6721</a>) at 1/50000 dilution

Predicted band size: 32 kDa

Observed band size: 32 kDa,50 kDa

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