Anti-ANP32E antibody [EPR29188-30]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ANP32E antibody [EPR29188-30] (AB324142)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling ANP32E with ab324142 at 1/200 (2.335 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil. The section was incubated with ab324142 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-ANP32E antibody [EPR29188-30] (AB324142)

Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling ANP32E with ab324142 at 1/200 (2.335 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human thyroid carcinoma. The section was incubated with ab324142 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-ANP32E antibody [EPR29188-30] (AB324142)

Immunohistochemical analysis of paraffin-embedded Mouse breast carcinoma tissue labeling ANP32E with ab324142 at 1/500 (0.934 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse breast carcinoma. The section was incubated with ab324142 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-ANP32E antibody [EPR29188-30] (AB324142)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling ANP32E with ab324142 at 1/500 (0.934 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebellum. The section was incubated with ab324142 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

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-ANP32E antibody [EPR29188-30] (AB324142)

Chromatin was prepared from MEF cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 ug of ab324142 [EPR29188-30]. 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-ANP32E antibody [EPR29188-30] (AB324142)

ANP32E was immunoprecipitated from 0.35 mg MEF (mouse embryo fibroblast) whole cell lysate with ab324142 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324142 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-ANP32E antibody [EPR29188-30] (ab324142) at 1/1000 dilution

Lane 1:

MEF (mouse embryo fibroblast) whole cell lysate() at 10 µg

Lane 2:

ab324142 IP in MEF (mouse embryo fibroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab324142 in MEF whole cell lysate

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: 31 kDa

false

Exposure time: 24s

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-ANP32E antibody [EPR29188-30] (AB324142)

Chromatin was prepared from MEF cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 ug of ab324142 [EPR29188-30]. 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-ANP32E antibody [EPR29188-30] (AB324142)

Chromatin was prepared from MEF cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 ug of ab324142 [EPR29188-30]. 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-ANP32E antibody [EPR29188-30] (AB324142)

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

Lanes 1-2 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross reactivity with human IgG at 1/2000 and lanes 3-6 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

Exposure time : Lanes 1-2 : 180 seconds; Lanes 3-6 : 26 seconds.

All lanes:

Western blot - Anti-ANP32E antibody [EPR29188-30] (ab324142) at 1/1000 dilution

Lane 1:

Human breast tissue lysate at 20 µg

Lane 2:

Human testis tissue lysate at 20 µg

Lane 3:

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

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

SR (human pleural effusion lymphoblast) whole cell lysate at 20 µg

Lane 6:

AsPC-1 (human Pancreas epithelial cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 2:

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

Lanes 3 - 6:

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: 31 kDa

false

• WB

Supplier Data

Western blot - Anti-ANP32E antibody [EPR29188-30] (AB324142)

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

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

All lanes:

Western blot - Anti-ANP32E antibody [EPR29188-30] (ab324142) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with siRNA specifically targeting ANP32E whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical 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 1/20000 dilution

Observed band size: 31 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-ANP32E antibody [EPR29188-30] (AB324142)

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

Exposure time : Lanes 1-4 : 37 seconds; Lanes 5-6 : 10 seconds.

All lanes:

Western blot - Anti-ANP32E antibody [EPR29188-30] (ab324142) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse breast tissue lysate at 20 µg

Lane 4:

MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 6:

C2C12 (mouse myoblast) 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 1/20000 dilution

Observed band size: 31 kDa

false

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-ANP32E antibody [EPR29188-30] (AB324142)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ MEF (mouse embryo fibroblast) cells and 5 ug of ab324142 [EPR29188-30]. 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-ANP32E antibody [EPR29188-30] (AB324142)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ MEF (mouse embryo fibroblast) cells and 5 ug of ab324142 [EPR29188-30]. 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-ANP32E antibody [EPR29188-30] (AB324142)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ MEF (mouse embryo fibroblast) cells and 5 ug of ab324142 [EPR29188-30]. 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.

• WB

Supplier Data

Western blot - Anti-ANP32E antibody [EPR29188-30] (AB324142)

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

This antibody does not cross-react with recombinant human ANP32A, ANP32B or ANP32D by western blot.

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

All lanes:

Western blot - Anti-ANP32E antibody [EPR29188-30] (ab324142) at 1/1000 dilution

Lane 1:

His-tagged human ANP32E recombinant protein at 10 ng

Lane 2:

His-tagged human ANP32A recombinant protein at 10 ng

Lane 3:

His-tagged human ANP32B recombinant protein at 10 ng

Lane 4:

His-tagged human ANP32D recombinant protein at 10 ng

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: 14 kDa,15 kDa

false

Exposure time: 26s