Anti-VTI1B antibody [EPR15506(B)]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VTI1B antibody [EPR15506(B)] (AB184170)

Immunofluorescent analysis of MCF7 cells (paraformaldehyde-fixed, 4%) labeling VTI1B with ab184170 at 1/50 dilution followed by Goat anti rabbit IgG (AlexaFluor^® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VTI1B antibody [EPR15506(B)] (AB184170)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma labeling VTI1B with ab184170 at 1 : 100 diluton and HRP polymer. Counterstained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-VTI1B antibody [EPR15506(B)] (AB184170)

Intracellular flow cytometric analysis of Hela cells (2% paraformaldehyde-fixed) labeling VTI1B with ab184170 at 1/170 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.

• WB

Lab

Western blot - Anti-VTI1B antibody [EPR15506(B)] (AB184170)

Lanes 1-3 : Merged signal (red and green). Green - ab184170 observed at 28 kDa. Red - loading control ab8245 observed at 36 kDa.

ab184170 Anti-VTI1B antibody [EPR15506(B)] was shown to specifically react with VTI1B in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265483 (knockout cell lysate ab257797) was used. Wild-type and VTI1B knockout samples were subjected to SDS-PAGE. ab184170 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

VTI1B knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human VTI1B knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-vti1b-knockout-hela-cell-line-ab265483'>ab265483</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 27 kDa

Observed band size: 28 kDa

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• WB

Supplier Data

Western blot - Anti-VTI1B antibody [EPR15506(B)] (AB184170)

All lanes:

Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170) at 1/10000 dilution

All lanes:

MCF7 cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 27 kDa

false

• WB

Supplier Data

Western blot - Anti-VTI1B antibody [EPR15506(B)] (AB184170)

All lanes:

Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170) at 1/2000 dilution

Lane 1:

MCF7 cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

Human fetal liver lysate at 20 µg

Lane 4:

Human fetal lung lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 27 kDa

false