Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (AB190357)

Intracellular flow cytometrical analysis of U937 cells labeling SERPINB1/PI2 with ab190357 at 1/210 (red) compared to a Rabbit monoclonal IgG as an isotope control (green). FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (AB190357)

Immunofluorescent analysis of 4% paraformaldehyde fixed and permeabilized with 0.1% Triton X-100 HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling SERPINB1/PI2 with ab190357 at 1/100 dilution (10 μg/ml), followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (2 μg/ml) (green). Confocal image showing cytoplasmic staining in HepG2 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). DAPI was used as nuclear counterstain.
The negative controls are as follows :
-ve control 1 : ab190357 at 1/100 dilution (10 μg/ml), followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (2 μg/ml).
-ve control 2 : AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (2 μg/ml).

• WB

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Western blot - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (AB190357)

All lanes:

Western blot - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (ab190357) at 1/5000 dilution

Lane 1:

HepG2 cell lysate at 20 µg

Lane 2:

A549 cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa

Observed band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (AB190357)

All lanes:

Western blot - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (ab190357) at 1/5000 dilution

Lane 1:

Human fetal kidney lysate at 20 µg

Lane 2:

Human fetal lung lysate at 20 µg

Lane 3:

Human fetal liver lysate at 20 µg

Lane 4:

Human stomach lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 43 kDa

Observed band size: 43 kDa

false

• WB

Lab

Western blot - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (AB190357)

Western blot : Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker ab190357 staining at 1/5000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 40 kDa in Wild-type A549 cell lysates with no signal observed at this size in SERPINB1 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SERPINB1/PI2 antibody [EPR13299] - Membrane Vesicle Marker (ab190357) at 1/5000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

SERPINB1 knockout A549 at 20 µg

Lane 3:

SK-OV-3 at 20 µg

Lane 4:

HT1080 at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 43 kDa

Observed band size: 40 kDa,37 kDa

false