Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling PPM1H with ab303536 at 1/500 dilution (1.082 µg/mL), followed by a ready to use Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Negative control : no staining on human liver.

The section was incubated with ab303536 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 Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling PPM1H with ab303536 at 1/500 dilution (1.082 µg/mL), followed by a ready to use Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Positive staining was observed on human cerebrum.

The section was incubated with ab303536 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 Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling PPM1H with ab303536 at 1/100 dilution (5.41 µg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green).

Confocal image showing positive staining on rat cerebrum.

The section was incubated with ab303536 for 60 mins at room temperature. The section was then mounted using Fluoromount^®. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 dilution (2 µg/mL).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum (fresh) tissue labeling PPM1H with ab303536 at 1/100 dilution (5.41 µg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green).

Confocal image showing positive staining on mouse cerebrum.

The section was incubated with ab303536 for 60 mins at room temperature. The section was then mounted using Fluoromount^®. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 dilution (2 µg/mL).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling PPM1H with ab303536 at 1/2000 dilution (0.265 µg/mL), followed by a ready to use Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Positive staining was observed on rat cerebrum.

The section was incubated with ab303536 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 Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling PPM1H with ab303536 at 1/2000 dilution (0.265 µg/mL), followed by a ready to use Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Positive staining was observed on mouse cerebrum.

The section was incubated with ab303536 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 Leica DS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• WB

Lab

Western blot - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

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

Lysates were freshly made and used immediately to minimize protein degradation.

All lanes:

Western blot - Anti-PPM1H antibody [EPR26028-53] (<a href='/en-us/products/primary-antibodies/ppm1h-antibody-epr26028-53-ab303536'>ab303536</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

SH-SY5Y (human neuroblastoma epithelial 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 1/100000 dilution

Observed band size: 56 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

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

Lysates were freshly made and used immediately to minimize protein degradation.

All lanes:

Western blot - Anti-PPM1H antibody [EPR26028-53] (<a href='/en-us/products/primary-antibodies/ppm1h-antibody-epr26028-53-ab303536'>ab303536</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte), whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate  at 20 µg

Lane 3:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 4:

NR8383 (rat alveolar macrophage), 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: 56 kDa,56 Da

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

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

Exposure time : Lane 1 : 48 seconds; Lane 2 : 3 minutes; Lane 3 : 125 seconds.

In lane 1, the lysates were stored at -80°C prior to Western Blotting. The band beneath the target band (56 kDa) is expected to be degradation products.

In lane 2-3, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

The blot of lane 3 was developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-PPM1H antibody [EPR26028-53] (<a href='/en-us/products/primary-antibodies/ppm1h-antibody-epr26028-53-ab303536'>ab303536</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

MCF7 whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial 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 1/100000 dilution

Observed band size: 56 kDa

false

• WB

Lab

Western blot - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

PPM1H was immunoprecipitated from 0.35 mg mouse brain tissue lysate with ab303536 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab303536 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

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

All lanes:

Western blot - Anti-PPM1H antibody [EPR26028-53] (<a href='/en-us/products/primary-antibodies/ppm1h-antibody-epr26028-53-ab303536'>ab303536</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/ppm1h-antibody-epr26028-53-ab303536'>ab303536</a> at 1/30 IP in Mouse brain tissue lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/ppm1h-antibody-epr26028-53-ab303536'>ab303536</a> in Mouse brain tissue lysate at 10 µg

Secondary

All lanes:

Western blot - 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: 56 kDa

false

Exposure time: 127s

• WB

Lab

Western blot - Anti-PPM1H antibody [EPR26028-53] - BSA and Azide free (AB303537)

This data was developed using ab303536, the same antibody clone in a different buffer formulation.

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

Lysates were freshly made and used immediately to minimize protein degradation

All lanes:

Western blot - Anti-PPM1H antibody [EPR26028-53] (<a href='/en-us/products/primary-antibodies/ppm1h-antibody-epr26028-53-ab303536'>ab303536</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse spleen 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: 56 kDa

false

Exposure time: 15s