COX IV Recombinant Rabbit Monoclonal Antibody [JJ09-05]
cat.: ET1701-63
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JJ09-05
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 20 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human COX IV aa 21-70 / 169.
Positive control: HeLa cell lysate, HepG2 cell lysate, mouse heart tissue lysate, rat heart tissue lysate, HepG2, MCF-7, human liver tissue, human liver carcinoma tissue, human colon carcinoma tissue, human kidney tissue, mouse colon tissue, mouse heart tissue.
Subcellular location: Mitochondrion inner membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:1,000-1:5,000
1:50-1:200
1:200
1:400-1:1,000
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P13073 Human | P19783 Mouse | P10888 Rat
Alternative names: AL024441 COX 4 COX IV 1 COX IV COX IV-1 Cox4 COX41_HUMAN Cox4a COX4B COX4I1 COX4I2 COX4L2 COXIV Cytochrome c oxidase polypeptide IV Cytochrome c oxidase subunit 4 isoform 1 mitochondrial Cytochrome c oxidase subunit 4 isoform 1, mitochondrial Cytochrome C Oxidase subunit IV Cytochrome c oxidase subunit IV isoform 1 Cytochrome c oxidase subunit IV isoform 2 (lung) Cytochrome c oxydase subunit 4 dJ857M17.2 MGC105470 MGC72016
Images
ET1701-63_1.jpg Fig1: Western blot analysis of COX IV on different lysates with Rabbit anti-COX IV antibody (ET1701-63) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: HepG2 cell lysate (20 µg/Lane)
Lane 3: Mouse heart tissue lysate (30 µg/Lane)
Lane 4: Rat heart tissue lysate (30 µg/Lane)

Predicted band size: 20 kDa
Observed band size: 15 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-63) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1701-63_2.jpg Fig2: ICC staining of COX IV in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-63, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-63_3.jpg Fig3: Immunocytochemistry analysis of MCF-7 cells labeling COX IV with Rabbit anti-COX IV antibody (ET1701-63) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-COX IV antibody (ET1701-63) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
ET1701-63_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue with Rabbit anti-COX IV antibody (ET1701-63) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-63) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-63_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-COX IV antibody (ET1701-63) at 1/800 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-63) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-63_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-COX IV antibody (ET1701-63) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-63) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-63_7.jpg Fig7: Flow cytometric analysis of COX IV was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-63, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1701-63_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-COX IV antibody (ET1701-63) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-63) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.