Anti-MDMX/MDM4 antibody [EPR30428-545]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling MDMX/MDM4 with ab325337 at 1/1000 (0.511 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat testis.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling MDMX/MDM4 with ab325337 at 1/1000 (0.511 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis (PMID : 26595814).

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

• IP

Lab

Immunoprecipitation - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

MDMX/MDM4 was immunoprecipitated from 0.35 mg Mouse testis tissue lysate with ab325337 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325337 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse testis tissue lysate
Lane 2 : ab325337 IP in Mouse testis tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325337 in mouse testis whole cell lysate

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

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-MDMX/MDM4 antibody [EPR30428-545] (ab325337) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 10 µg

Lane 2:

ab325337 at 1/30 IP in Mouse testis tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325337 in mouse testis whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 65 kDa,55 kDa

false

Exposure time: 180s

• IP

Lab

Immunoprecipitation - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

MDMX/MDM4 was immunoprecipitated from 0.35 mg Rat testis tissue lysate with ab325337 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325337 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat testis tissue lysate
Lane 2 : ab325337 IP in Rat testis tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325337 in rat testis whole cell lysate

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

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-MDMX/MDM4 antibody [EPR30428-545] (ab325337) at 1/1000 dilution

Lane 1:

Rat testis tissue lysate at 10 µg

Lane 2:

ab325337 at 1/30 IP in Rat testis tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325337 in rat testis whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 65 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

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

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MDMX/MDM4 antibody [EPR30428-545] (ab325337) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma t lymphocyte) fresh whole cell lysate at 20 µg

Lane 2:

EL4 (mouse lymphoma t lymphocyte) frozen 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: 65 kDa,55 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

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

Negative control : colon (PMID : 9226370), skeletal muscle (PMID : 26595814).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MDMX/MDM4 antibody [EPR30428-545] (ab325337) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 3:

Mouse colon 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 1/20000 dilution

Observed band size: 65 kDa,55 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

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

Negative control : colon (PMID : 9226370).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-MDMX/MDM4 antibody [EPR30428-545] (ab325337) at 1/1000 dilution

Lane 1:

Rat testis tissue lysate at 20 µg

Lane 2:

Rat colon 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 1/20000 dilution

Observed band size: 65 kDa,55 kDa,36 kDa

false

Exposure time: 180s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MDMX/MDM4 with ab325337 at 1/1000 (0.511 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse skeletal muscle (PMID : 26595814).

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling MDMX/MDM4 with ab325337 at 1/1000 (0.511 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse colon (PMID : 26595814).

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling MDMX/MDM4 with ab325337 at 1/1000 (0.511 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on rat skeletal muscle.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling MDMX/MDM4 with ab325337 at 1/1000 (0.511 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on rat colon.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDMX/MDM4 antibody [EPR30428-545] (AB325337)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a MDM4 expression vector containing a Myc-His tag. (B) HEK-293T transfected with empty vector containing a Myc-His tag. tissue labeling MDMX/MDM4 with ab325337 at 1/6000 (0.085 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a MDM4 expression vector containing a Myc-His tag, no staining on (B) HEK-293T transfected with empty vector containing a Myc-His tag.

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