Anti-AP2 gamma/TFAP2C antibody [EPR29851-13]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling AP2 gamma/TFAP2C with ab324045 at 1/500 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat kidney. The section was incubated with ab324045 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling AP2 gamma/TFAP2C with ab324045 at 1/500 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse kidney (PMID : 9765260). The section was incubated with ab324045 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse placenta tissue staining PLGF with ab322267 at a 1/2000 dilution, ab324045 anti-AP2 gamma/TFAP2C used at 1/500 dilution and ab182981 anti-CD31 used at a 1/5000 dilution.

Panel A : merged staining of anti-PLGF (green; Opal™520), anti-AP2 gamma/TFAP2C (magenta; Opal™690) and anti-CD31 (gray; Opal™570) on mouse placenta.

Panel B : anti-PLGF showed positive staining in mouse placenta.

Panel C : anti-AP2 gamma/TFAP2C staining trophoblast stem cells in mouse placenta.

Panel D : anti-CD31 staining endothelium in mouse placenta.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322267, ab324045 and ab182981 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse placenta tissue staining LOX 1 with ab317689 at a 1/2000 dilution, ab324045 anti-AP2 gamma used at 1/500 dilution and ab182981 anti-CD31 used at a 1/5000 dilution.

Panel A : merged staining of anti-LOX 1 (green; Opal™520), anti-AP2 gamma (magenta; Opal™690) and anti-CD31 (gray; Opal™570) on mouse placenta.

Panel B : anti-LOX 1 staining trophoblast in mouse placenta.

Panel C : ant-AP2 gamma staining trophoblast stem cell in mouse placenta.

Panel D : ant-CD31 staining endothelium in mouse placenta.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317689, ab324045 and ab182981 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse placenta tissue staining LOX 1 with ab317689 at a 1/2000 dilution, ab324045 anti-AP2 gamma used at 1/500 dilution and ab182981 anti-CD31 used at a 1/5000 dilution.

Panel A : merged staining of anti-LOX 1 (green; Opal™520), anti-AP2 gamma (magenta; Opal™690) and anti-CD31 (gray; Opal™570) on mouse placenta.

Panel B : anti-LOX 1 staining trophoblast in mouse placenta.

Panel C : ant-AP2 gamma staining trophoblast stem cell in mouse placenta.

Panel D : ant-CD31 staining endothelium in mouse placenta.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317689, ab324045 and ab182981 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse placenta tissue staining LOX 1 with ab317689 at a 1/2000 dilution, ab324045 anti-AP2 gamma used at 1/500 dilution and ab320823 anti-Trophoblast specific protein alpha used at a 1/1000 dilution.

Panel A : merged staining of anti-LOX 1 (green; Opal™520), anti-AP2 gamma (magenta; Opal™690) and anti-Trophoblast specific protein alpha (gray; Opal™570) on mouse placenta.

Panel B : anti-LOX 1 staining trophoblast in mouse placenta.

Panel C : ant-AP2 gamma staining trophoblast stem cell in mouse placenta.

Panel D : ant-Trophoblast specific protein alpha staining junctional zone in mouse placenta.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab317689, ab324045 and ab320823 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Immunohistochemical analysis of paraffin-embedded Mouse placenta tissue labeling AP2 gamma/TFAP2C with ab324045 at 1/500 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse placenta (PMID : 9765260). The section was incubated with ab324045 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat-1 (rat embryonic fibroblast) cells labelling AP2 gamma/TFAP2C with ab324045 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 (2ug/ml) dilution (Green).

Confocal image showing nuclear staining in Rat-1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled 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).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) cells labelling AP2 gamma/TFAP2C with ab324045 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 (2ug/ml) dilution (Green).

Confocal image showing nuclear staining in ES-D3 [D3] cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Low expression : MEF (PMID : 32587258)

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).

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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Immunohistochemical analysis of paraffin-embedded Rat placenta tissue labeling AP2 gamma/TFAP2C with ab324045 at 1/500 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat placenta. The section was incubated with ab324045 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Chromatin was prepared from ES-D3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5x10⁶ cells and 4 µg of ab324045 [EPR29851-13]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Supplier Data

Immunoprecipitation - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

AP2 gamma/TFAP2C was immunoprecipitated from 0.35 mg ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate with ab324045 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324045 at 1000 dilution.

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

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

All lanes:

Immunoprecipitation - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (ab324045) at 1/1000 dilution

Lane 1:

ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate at 10 µg

Lane 2:

ab324045 at 1/30 IP in ES-D3 [D3] whole cell 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 ab324045 in ES-D3 [D3] whole cell lysate at 10 µg

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: 49 kDa,51 kDa

false

Exposure time: 180s

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Chromatin was prepared from ES-D3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5x10⁶ cells and 4 µg of ab324045 [EPR29851-13]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

Chromatin was prepared from ES-D3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5x10⁶ cells and 4 µg of ab324045 [EPR29851-13]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• WB

Supplier Data

Western blot - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

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

Low expression : MEF (PMID : 32587258).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32587258, PMID : 29695788).

In lanes 3-4, the lysates were stored at -80degC prior to Western Blotting. The bands beneath the target band (51 kDa) are likely to be degradation products. In lanes 1-2, To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the bands higher than 75kDa are unknown.

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

All lanes:

Western blot - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (ab324045) at 1/1000 dilution

Lane 1:

ES-D3 D3  (mouse blastocyst-derived embryonic stem cell) fresh whole cell lysate at 50 µg

Lane 2:

MEF (mouse embryo fibroblast) fresh whole cell lysate at 50 µg

Lane 3:

ES-D3 D3  (mouse blastocyst-derived embryonic stem cell) frozen whole cell lysate at 50 µg

Lane 4:

MEF (mouse embryo fibroblast) frozen whole cell lysate at 50 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: 49 kDa,51 kDa,36 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

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

Negative control : kidney (PMID : 9765260).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32587258, PMID : 29695788).

The identity of the bands higher than 75kDa are unknown.

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

All lanes:

Western blot - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (ab324045) at 1/1000 dilution

Lane 1:

Rat-1 (rat embryonic fibroblast) whole cell lysate at 50 µg

Lane 2:

Mouse placenta tissue lysate at 50 µg

Lane 3:

Mouse kidney tissue lysate at 50 µg

Lane 4:

Rat placenta tissue lysate at 50 µg

Lane 5:

Rat kidney tissue lysate at 50 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: 49 kDa,51 kDa,36 kDa

false

Exposure time: 180s

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) cells and 5 µg of ab324045 [EPR29851-13]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) cells and 5 µg of ab324045 [EPR29851-13]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-AP2 gamma/TFAP2C antibody [EPR29851-13] (AB324045)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) cells and 5 µg of ab324045 [EPR29851-13]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.