Anti-MTCO1 antibody [1D6E1A8]

• WB

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Western blot - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MTCO1 antibody [1D6E1A8] (ab14705) 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

Lane 3:

MCF7(human breast adenocarcinoma epithelial cell) non-mitochondrial fraction at 20 µg

Lane 4:

MCF7 mitochondrial fraction at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 35 kDa

false

• Flow Cyt

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Flow Cytometry - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Flow cytometric analysis of Human HeLa cells labelling MTCO1 with ab14705 at 1/500 dilution (Right) compared with an isotype control (Left). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at was used as the secondary antibody. Gated on viable cells.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Human cervical adenocarcinoma epithelial cells labeling MTCO1 with ab14705 at 1/100 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution. Confocal image showing cytoplasmic and membranous staining in subsets of Human cervical adenocarcinoma epithelial cells. ab186735 Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker was used to counterstain tubulin at 1/400 dilution. The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling MTCO1 with ab14705 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Human kidney. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. 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 Solution2) for 20 mins. Counterstained with hematoxylin.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

This image was generated using a previous batch manufactured using hybridoma production method.

250 μm human cerebellar sections from control individuals and a patient with mitochondrial disease underwent passive clearing at 37 °C for 2 or 4 weeks.

The quality of immunofluorescent staining is determined by duration of passive clearing; 2 weeks of passive clearing produced minimal labelling of the white matter in the granule cell layer (NF-H; green; 488 nm and MBP; red, 546 nm) with an absence of labelling of mitochondria (MTCO1 (COXI) (ab14705, 1/100); purple; 647 nm; Extending passive clearing to 4 weeks improved the quality of stain with identifiable Purkinje cells and their axons (NF-H, green; 488 nm) and their myelin sheaths (MBP; red, 546 nm) and mitochondria (MTCO1 (COXI) (ab14705, 1/100); purple; 647 nm.

Image from Phillips J et al., Sci Rep. 2016 May 16;6:26013. Fig 3b doi: 10.1038/srep26013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labelling MTCO1 with ab14705 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Rat colon. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. 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 Solution2) for 20 mins. Counterstained with hematoxylin.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Flow cytometric analysis of Neuro-2a cells labelling MTCO1 with ab14705 at 1/500 dilution (Right) compared with an isotype control (Left). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at was used as the secondary antibody. Gated on viable cells.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse neuroblastoma neuroblast cells labeling MTCO1 with ab14705 at 1/100 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution. Confocal image showing cytoplasmic and membranous staining in subsets of Mouse neuroblastoma neuroblast cells. ab186735 Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker was used to counterstain tubulin at 1/400 dilution. The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling MTCO1 with ab14705 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Mouse colon. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. 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 Solution2) for 20 mins. Counterstained with hematoxylin.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat glial tumor cells labeling MTCO1 with ab14705 at 1/100 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution. Confocal image showing cytoplasmic and membranous staining in subsets of Rat glial tumor cells. ab186735 Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker was used to counterstain tubulin at 1/400 dilution. The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MTCO1 antibody [1D6E1A8] (ab14705) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 5:

Human skeletal muscle tissue lysate at 20 µg

Lane 6:

Rat heart tissue lysate at 20 µg

Lane 7:

Rat liver tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 35 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MTCO1 antibody [1D6E1A8] (AB14705)

Immunofluorscent analysis of NIH 3T3, COS7, H9C2 and HeLa cells using ab170192 at 1μg/ml (top); no primary antibody negative control (middle), and ab14705 as a positive control (bottom). An Alexa Fluor® 488 conjugate with isotype specificity to the mouse antibody was used as a secondary antibody. The isotype control at 1 ug/mL shows no higher signal than the no primary negative control.