Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free (AB314125)

This data was developed using ab314124, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling DOM3Z with ab314124 at 1/50 (10.1 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 mainly nuclear staining in HeLa cell line. 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 2.5ug/ml dilution (Red). 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 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free (AB314125)

This data was developed using ab314124, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling DOM3Z with ab314124 at 1/50 dilution (1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free (AB314125)

This data was developed using ab314124, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling DOM3Z with ab314124 at 1/1000 (0.505 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Nuclear staining on mouse testis. The section was incubated with ab314124 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free (AB314125)

This data was developed using ab314124, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling DOM3Z with ab314124 at 1/1000 (0.505 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Nuclear staining on rat testis. The section was incubated with ab314124 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).

• WB

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Western blot - Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free (AB314125)

This data was developed using ab314124, the same antibody clone in a different buffer formulation.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The identity of the bands between 25 kDa and 37 kDa are unknown.

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

All lanes:

Western blot - Anti-DOM3Z antibody [EPR27511-140] (<a href='/en-us/products/primary-antibodies/dom3z-antibody-epr27511-140-ab314124'>ab314124</a>) at 1/1000 dilution

Lane 1:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

Raji (human Burkitts lymphoma B lymphocyte) 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: 45 kDa

true

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free (AB314125)

This data was developed using ab314124, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-DOM3Z antibody [EPR27511-140] (<a href='/en-us/products/primary-antibodies/dom3z-antibody-epr27511-140-ab314124'>ab314124</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa transfected with siRNA specifically targeting DOM3Z 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: 45 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-DOM3Z antibody [EPR27511-140] - BSA and Azide free (AB314125)

This data was developed using ab314124, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the bands between 20 kDa and 37 kDa are unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-DOM3Z antibody [EPR27511-140] (<a href='/en-us/products/primary-antibodies/dom3z-antibody-epr27511-140-ab314124'>ab314124</a>) at 1/1000 dilution

Lane 1:

Mouse pancreas tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Lane 4:

Rat pancreas 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: 45 kDa

false

Exposure time: 180s