Antibodies, Proteins, Kits and Reagents for Life Science

338 390 ₸ Product size

100 µl 338 390 ₸

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Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR9442(ABC)] to COX IV - Mitochondrial Loading Control
Suitable for: ICC/IF, IP, Flow Cyt, IHC-P, WB
Reacts with: Mouse, Rat, Human

Product name

Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control
See all COX IV primary antibodies
Description

Rabbit monoclonal [EPR9442(ABC)] to COX IV - Mitochondrial Loading Control
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal heart lysate; HepG2 whole cell lysate; Mouse and rat heart lysates. IHC-P: Human hepatocellular carcinoma, Human cervix carcinoma, mouse kidney and rat cardiac muscle tissues. ICC/IF: HeLa and HepG2 cells. Flow Cyt: MCF7 cells. IP: Human fetal heart whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR9442(ABC)
Isotype

IgG
Research areas

Western blot - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

All lanes : Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554) at 1/2000 dilution

Lane 1 : Human fetal heart lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 20 kDa
Observed band size: 17 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling COX IV with ab202554 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Cytoplasmic staining on HepG2 cells is observed.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab202554 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling COX IV with ab202554 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Cytoplasmic staining on HeLa cells is observed.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab202554 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Western blot - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

All lanes : Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554) at 1/10000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Rat heart lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 20 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling COX IV with ab202554 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Cytoplasmic staining on Human hepatocellular carcinoma tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling COX IV with ab202554 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Cytoplasmic staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling COX IV with ab202554 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Cytoplasmic staining on mouse kidney tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling COX IV with ab202554 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Cytoplasmic staining on Human cervix carcinoma tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

Flow cytometric analysis of 2% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling COX IV with ab202554 at 1/20 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

COX IV was immunoprecipitated from 1mg of Human fetal heart whole cell lysate with ab202554 at 1/20 dilution.

Western blot was performed from the immunoprecipitate using ab202554 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.

Lane 1: Human fetal heart whole cell lysate 10 µg (Input).

Lane 2: ab202554 IP in Human fetal heart whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202554 in Human fetal heart whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 seconds.
Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554)

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