Anti-Total OXPHOS Antibody Cocktail antibody [RM1134]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (AB317270)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Total OXPHOS Antibody Cocktail with ab317270 at 1/500 dilution (0.1ug)/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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (AB317270)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Total OXPHOS Antibody Cocktail with ab317270 at 1/2000 (0.246 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast carcinoma. The section was incubated with ab317270 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

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (AB317270)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Total OXPHOS Antibody Cocktail with ab317270 at 1/500 (0.984 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 HepG2 cell line (shown in green). The counter stain was observed in magenta. Nuclear DNA was labeled 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 1/1000 1ug/ml dilution (Magenta). 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (AB317270)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Total OXPHOS Antibody Cocktail with ab317270 at 1/2000 (0.246 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver. The section was incubated with ab317270 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

Supplier Data

Western blot - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (AB317270)

All lanes:

Western blot - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (ab317270) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 18 kDa,22 kDa,29 kDa,48 kDa,54 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (AB317270)

Exposure time : Lanes 1-3 : 15 seconds; Lanes 4-5 : 37 seconds

All lanes:

Western blot - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (ab317270) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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

Lane 5:

MCF7 (human breast 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 1/100000 dilution

Observed band size: 18 kDa,22 kDa,29 kDa,48 kDa,54 kDa

false

• WB

Supplier Data

Western blot - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (AB317270)

In Western blot, Anti-Alpha-synuclein antibody [EPR20535] (ab212184) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Total OXPHOS Antibody Cocktail antibody [RM1134] (ab317270) at 1/1000 dilution

Lane 1:

MCF7 non-mitochondrial fraction at 20 µg

Lane 2:

MCF7 mitochondria fraction 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 1/100000 dilution

Observed band size: 18 kDa,22 kDa,29 kDa,48 kDa,54 kDa,36 kDa,17 kDa

false

Exposure time: 8s