Anti-PADI4 / PAD4 antibody [EPR26687-49]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling PADI4 / PAD4 with ab315269 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human cerebrum (PMID : 12833157).
The section was incubated with ab315269 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-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling PADI4 / PAD4 with ab315269 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on granulocytes in human spleen.
The section was incubated with ab315269 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-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling PADI4 / PAD4 with ab315269 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on granulocytes in human liver.
The section was incubated with ab315269 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-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling PADI4 / PAD4 with ab315269 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on granulocytes in human lung.
The section was incubated with ab315269 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-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HL-60 (human acute promyelocytic leukemia promyeloblast) cells labelling PADI4 / PAD4 with ab315269 at 1/500 (1.026 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 HL-60 cells treated with 1.25% DMSO for 5 days and showing no staining in untreated HL-60 cells.
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 (Red). 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-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HL-60 (human acute promyelocytic leukemia promyeloblast) treated with 1.25% DMSO for 5days) (Red) / untreated HL-60 (Green) cells labelling PADI4 / PAD4 with ab315269 at 1/5000 dilution (0.01 ug)/Red and Green 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.

• WB

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Western blot - Anti-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

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

The expression of PAD is upregulated in response to DMSO treatment (PMID : 10488123).

Lysates were freshly made and used 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-PADI4 / PAD4 antibody [EPR26687-49] (ab315269) at 1/1000 dilution

Lane 1:

Untreated HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

Lane 2:

HL-60 treated with 1.25% DMSO for 5 days 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: 74 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-PADI4 / PAD4 antibody [EPR26687-49] (AB315269)

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

This antibody does not cross-react with human PADI1, PADI2, PADI3 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-PADI4 / PAD4 antibody [EPR26687-49] (ab315269) 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 PADI4 expression vector containing a myc-His-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 PADI2 expression vector containing a GFP-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: 75 kDa,36 kDa

false

Exposure time: 6s