Anti-CHCHD2 antibody [EPR29117-175]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHCHD2 antibody [EPR29117-175] (AB322258)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CHCHD2 with ab322258 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human colon.
The section was incubated with ab322258 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-CHCHD2 antibody [EPR29117-175] (AB322258)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CHCHD2 with ab322258 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining in human liver (PMID : 31839816).
The section was incubated with ab322258 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-CHCHD2 antibody [EPR29117-175] (AB322258)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling CHCHD2 with ab322258 at 1/50 (10.08 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 mitochondrial staining in HeLa cells (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).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab322258 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. -ve control 2 : anti-COX IV mouse monoclonal antibody at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHCHD2 antibody [EPR29117-175] (AB322258)

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling CHCHD2 with ab322258 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human astrocytoma.
The section was incubated with ab322258 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-CHCHD2 antibody [EPR29117-175] (AB322258)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling CHCHD2 with ab322258 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human cerebrum.
The section was incubated with ab322258 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-CHCHD2 antibody [EPR29117-175] (AB322258)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling CHCHD2 with ab322258 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human colon cancer.
The section was incubated with ab322258 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

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Western blot - Anti-CHCHD2 antibody [EPR29117-175] (AB322258)

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

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-CHCHD2 antibody [EPR29117-175] (ab322258) at 1/1000 dilution

All lanes:

Human liver cancer tissue lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 17 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CHCHD2 antibody [EPR29117-175] (AB322258)

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-CHCHD2 antibody [EPR29117-175] (ab322258) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting CHCHD2 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: 17 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CHCHD2 antibody [EPR29117-175] (AB322258)

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

All lanes:

Western blot - Anti-CHCHD2 antibody [EPR29117-175] (ab322258) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) whole lysate at 20 µg

Lane 4:

LNCaP (human prostate carcinoma epithelial cell) whole 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: 17 kDa

false

Exposure time: 37s

• Dot

Supplier Data

Dot Blot - Anti-CHCHD2 antibody [EPR29117-175] (AB322258)

Dot blot analysis of CHCHD2 using ab322258 at 1 : 1000 (0.504 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : No tagged human CHCHD2 fragment
Lane2 : His-tagged human CHCHD10 fragment

Exposure time : 26 seconds.

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

This antibody does not cross-react with human CHCHD10.

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

All lanes:

Dot Blot - Anti-CHCHD2 antibody [EPR29117-175] (ab322258) at 1/1000 dilution

Lane 1:

No tagged human CHCHD2 fragment

Lanes 1 - 2:

His-tagged human CHCHD10 fragment

Secondary

All lanes:

Dot 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

false

Exposure time: 26s