Anti-VAPB antibody [EPR27026-59]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VAPB antibody [EPR27026-59] (AB324047)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling VAPB with ab324047 at 1/50 (9.76 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

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

Anti-KDEL mouse monoclonal antibody was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab324047 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. -ve control 2 : Anti-KDEL mouse monoclonal antibody at 1/200 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-VAPB antibody [EPR27026-59] (AB324047)

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

All lanes:

Western blot - Anti-VAPB antibody [EPR27026-59] (ab324047) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocelullar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Human testis tisse lysate at 20 µg

Lane 4:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 27 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-VAPB antibody [EPR27026-59] (AB324047)

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

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

All lanes:

Western blot - Anti-VAPB antibody [EPR27026-59] (ab324047) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting VAPB whole cell 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: 27 kDa,36 kDa

false

Exposure time: 26s

• Dot

Supplier Data

Dot Blot - Anti-VAPB antibody [EPR27026-59] (AB324047)

Dot blot analysis of VAPB using ab324047 at 1 : 1000 (0.488 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : His-tagged human VAPB recombinant protein
Lane 2 : His-tagged human VAPA recombinant protein

Exposure time : 180 seconds.

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

This antibody does not cross-react with human VAPA.

In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-VAPB antibody [EPR27026-59] (ab324047) at 1/1000 dilution

Lane 1:

His-tagged human VAPB recombinant protein

Lane 2:

His-tagged human VAPA recombinant protein

Secondary

All lanes:

Dot 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

false

Exposure time: 180s