Anti-PML Protein antibody [EPR27157-75]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR27157-75] (AB317018)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling PML Protein with ab317018 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on mouse cerebrum.
The section was incubated with ab317018 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-PML Protein antibody [EPR27157-75] (AB317018)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling PML Protein with ab317018 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on rat cerebrum.
The section was incubated with ab317018 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-PML Protein antibody [EPR27157-75] (AB317018)

Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling PML Protein with ab317018 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung cancer.
The section was incubated with ab317018 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-PML Protein antibody [EPR27157-75] (AB317018)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling PML Protein with ab317018 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen.
The section was incubated with ab317018 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-PML Protein antibody [EPR27157-75] (AB317018)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PML Protein with ab317018 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab317018 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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PML Protein antibody [EPR27157-75] (AB317018)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling PML Protein with ab317018 at 1/500 (0.98 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 nuclear staining in F9 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
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 (Magenta). 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-PML Protein antibody [EPR27157-75] (AB317018)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling PML Protein with ab317018 at 1/500 dilution (0.1 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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-PML Protein antibody [EPR27157-75] (AB317018)

PML Protein was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate with ab317018 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317018 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate
Lane 2 : ab317018 IP in F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317018 in F9 whole cell lysate

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 28486108).

All lanes:

Immunoprecipitation - Anti-PML Protein antibody [EPR27157-75] (ab317018) at 1/30 dilution

All lanes:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

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

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-PML Protein antibody [EPR27157-75] (AB317018)

Lane 5 was freshly made and used for Western Blotting immediately to minimize protein degradation.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 28486108).

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

All lanes:

Western blot - Anti-PML Protein antibody [EPR27157-75] (ab317018) at 1/1000 dilution

Lane 1:

MEF (mouse embryo fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

MEF transfected with siRNA specifically targeting PML Protein whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 5:

F9 whole fresh cell lysate at 20 µg with 5% NFDM/TBST

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: 110 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PML Protein antibody [EPR27157-75] (AB317018)

Low expression : brain

The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-PML Protein antibody [EPR27157-75] (ab317018) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg with 5% NFDM/TBST

Lane 2:

Mouse brain tissue lysate at 20 µg with 5% NFDM/TBST

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: 110 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-PML Protein antibody [EPR27157-75] (AB317018)

Low expression : brain

The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-PML Protein antibody [EPR27157-75] (ab317018) at 1/1000 dilution

Lane 1:

Rat spleen tissue lysate at 20 µg with 5% NFDM/TBST

Lane 2:

Rat brain tissue lysate at 20 µg with 5% NFDM/TBST

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: 110 kDa,15 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-PML Protein antibody [EPR27157-75] (AB317018)

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

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

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

All lanes:

Western blot - Anti-PML Protein antibody [EPR27157-75] (ab317018) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human PML protein expression vector containing a myc-His-tag®, 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: 110 kDa,15 kDa

false

Exposure time: 6s