Anti-PADI2 / PAD2 antibody [EPR26940-85]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon (PMID : 28403548; PMID : 31268166). The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-BR-3 (human breast adenocarcinoma epithelial cell) cells labelling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in SK-BR-3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : MCF-7 (PMID : 23110523).
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 2ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on human liver (PMID : 23026755). The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen (PMID : 28993780). The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum. The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on rat liver. The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum. The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen. The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on mouse liver (PMID : 30071078; PMID : 23026755). The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PADI2 / PAD2 with ab322197 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen (PMID : 23026755). The section was incubated with ab322197 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-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

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

Low expression : liver, lung (PMID : 23026755).

Lanes 1-4 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and Lanes 4-9 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000.

The identity of the bands beneath 50 kDa (in lanes 1-2 ) are unknown.

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

All lanes:

Western blot - Anti-PADI2 / PAD2 antibody [EPR26940-85] (ab322197) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human lung tissue lysate at 20 µg

Lane 5:

Mouse colon tissue lysate at 20 µg

Lane 6:

Mouse brain tissue lysate at 20 µg

Lane 7:

Mouse liver tissue lysate at 20 µg

Lane 8:

Rat brain tissue lysate at 20 µg

Lane 9:

Rat liver tissue lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 4 - 9:

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: 76 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

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

Low expression : MCF7 (PMID : 23110523).

The expression profile is consistent with what has been described in the literature (PMID : 23110523).

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

Exposure time : Lane 1 : 180 seconds; Lanes 2-4 : 26 seconds.

All lanes:

Western blot - Anti-PADI2 / PAD2 antibody [EPR26940-85] (ab322197) at 1/1000 dilution

Lane 1:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast adenocarcinoma 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: 76 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-PADI2 / PAD2 antibody [EPR26940-85] (AB322197)

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

This antibody does not cross-react with human PADI1, PADI3, PADI4 and PADI6.

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

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

All lanes:

Western blot - Anti-PADI2 / PAD2 antibody [EPR26940-85] (ab322197) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human PADI2 expression vector containing a GFP-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human PADI1 expression vector containing a GFP-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human PADI3 expression vector containing a GFP-tag, whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a human PADI4 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 6:

293T cells transfected with a human PADI6 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,36 kDa,75 kDa

false

Exposure time: 6s