Anti-NEDD4-2 antibody [EPR29028-16]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma and adjacent stomach tissue labeling NEDD4-2 with ab324872 at 1/200 (2.465 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining observed in adjacent human stomach tissue, with reduced expression of NEDD4L in gastric carcinoma tissue (PMID : 31673244).
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

Immunohistochemical analysis of paraffin-embedded (A) PC-3 (human prostate adenocarcinoma epithelial cell) cell pellet; (B) SK-BR-3 (human breast cancer cell) cell pellet labeling NEDD4-2 with ab324872 at 1/1000 (0.493 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) PC-3 cell pellet, weak staining on (B) SK-BR-3 cell pellet (PMID : 14615060).
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling NEDD4-2 with ab324872 at 1/200 (2.465 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

Immunohistochemical analysis of paraffin-embedded Human clear cell carcinoma of kidney and adjacent kidney tissue labeling NEDD4-2 with ab324872 at 1/200 (2.465 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining observed in adjacent human kidney tissue, with reduced expression of NEDD4L in clear cell carcinoma of kidney tissue (PMID : 34539897).
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling NEDD4-2 with ab324872 at 1/200 (2.465 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse colon.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NEDD4-2 with ab324872 at 1/200 (2.465 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

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

Low expression : SK-BR-3, T-47D (PMID : 14615060).

The identity of the bands lower than 75 kDa are unknown.

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

Exposure time : Lanes 1-5 : 48 seconds; Lane 6 : 15 seconds.

All lanes:

Western blot - Anti-NEDD4-2 antibody [EPR29028-16] (ab324872) at 1/1000 dilution

Lane 1:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 5:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

SK-MEL-28 (human malignant melanoma cell ) 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 1/20000 dilution

Observed band size: 110 kDa,135 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

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

The identity of the lower MW band at approximately 12 kDa is unknown.

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

All lanes:

Western blot - Anti-NEDD4-2 antibody [EPR29028-16] (ab324872) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

PC-3 transfected with siRNA specifically targeting NEDD4-2 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 1/20000 dilution

Observed band size: 110 kDa,135 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-NEDD4-2 antibody [EPR29028-16] (AB324872)

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

The identity of the bands lower than 75 kDa are unknown.

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

Exposure time : Lanes 1-2 : 180 seconds; Lane 3 : 48 seconds; Lanes 4-5 : 180 seconds.

All lanes:

Western blot - Anti-NEDD4-2 antibody [EPR29028-16] (ab324872) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

C2C12 (mouse myoblast) 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 1/20000 dilution

Observed band size: 110 kDa,135 kDa,36 kDa

false