Anti-CHCHD10 antibody [EPR26966-33]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% Methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling CHCHD10 with ab308510 at 1/50 (10.24 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 showing mitochondrial staining in HeLa cell line. 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 1/50 0.5 ug/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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling CHCHD10 with ab308510 at 1/500 (1.024 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human liver.The section was incubated with ab308510 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular tissue labelling CHCHD10 with ab308510 at 1/500 (1.024 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human hepatocellular carcinoma.The section was incubated with ab308510 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling CHCHD10 with ab308510 at 1/500 (1.024 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human spleen (PMID : 24934289).The section was incubated with ab308510 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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling CHCHD10 with ab308510 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

CHCHD10 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab308510 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308510 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab308510 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308510 in HeLa whole cell lysate

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

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-CHCHD10 antibody [EPR26966-33] (ab308510) at 1/1000 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) 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: 18 kDa

false

Exposure time: 8s

• WB

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Western blot - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Anti-SPG7 (ab305255) staining at 1/200000 dilution. Exposure time : 59 seconds.

All lanes:

Western blot - Anti-CHCHD10 antibody [EPR26966-33] (ab308510) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) non-mitochondrial fraction at 20 µg

Lane 2:

HeLa mitochondrial fraction at 20 µg

Secondary

All lanes:

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

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

Exposure time: 59s

• WB

Lab

Western blot - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Western blot : Rabbit Monoclonal[EPR26966-33] to CHCHD10 ab308510 staining at 0.1 µg/mL, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 18 kDa in Wild-type A549 cell lysates with no signal observed at this size in CHCHD10 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-CHCHD10 antibody [EPR26966-33] (ab308510) at 0.1 µg/mL

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

CHCHD10 knockout A549 at 20 µg

Lane 3:

LNCaP at 20 µg

Lane 4:

U-2 OS at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

• WB

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Western blot - Anti-CHCHD10 antibody [EPR26966-33] (AB308510)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds.

All lanes:

Rat-1 (rat embryonic fibroblast) whole cell lysate at 20 µg

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

Exposure time: 180s