Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [3]
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- Product number
- 14-9775-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- COX4 Monoclonal Antibody (IB52C31H10), eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The IB52C31H10 monoclonal antibody recognizes human COX IV. COX IV, also known as cytochrome c oxidase (COX) or Complex IV, is the terminal complex of the electron transport chain. COX IV is encoded by nuclear DNA and localized to the inner mitochondrial membrane. COX IV functions in energy metabolism and shuttles electrons from cytochrome c to oxygen. Dysfunction in COX IV is associated with increased production of reactive oxygen species and cellular toxicity. Genetic defects in COX IV results in severe, often fatal metabolic disorders that primarily affect tissues with high energy demands such as heart, brain, and muscle. Applications Reported: This IB52C31H10 antibody has been reported for use in immunoprecipitation, western blotting, immunohistochemical staining of frozen tissue sections, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, microscopy, ELISA, and immunocytochemistry. Applications Tested: This IB52C31H10 antibody has been tested by immunocytochemistry of methanol-fixed human cells and can be used at less than or equal to 1 µg/mL. The IB52C31H10 antibody has also been tested by immunohistochemistry of formalin-fixed paraffin embedded tissue using low pH antigen retrieval and can be used at less than or equal to 1 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- IB52C31H10
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- 4° C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of COX4 was achieved by transfecting Hep G2 with COX4 specific siRNAs (Silencer® select Product # S3386, S3387). Western blot analysis (Fig. a) was performed using Whole cell extracts from the COX4 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with COX4 Monoclonal Antibody (IB52C31H10), eBioscience™ (Product # 14-9775-82, 1 µg/mL concentration) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to COX4.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-COX4 Monoclonal Antibody (IB52C31H10), eBioscience™(Product # 14-9775-82) and a 15kDa band corresponding to COX4 was observed across the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), LNCaP (Lane 3), A-431 (Lane 4), Rat Heart (Lane 5), Mouse Liver (Lane 6) and Mouse Heart (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody at a concentration of 1 µg/mL and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry of fixed HeLa cells stained with 1 µg/mL of Mouse IgG1 K Isotype Control Purified (left) or 1 µg/mL of Anti-Human COX IV Purified (right) followed by F (ab')2 Anti-Mouse IgG eFluor® 570.Nuclei are stained with DAPI.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry of fixed HeLa cells stained with 1 µg/mL of Mouse IgG1 K Isotype Control Purified (left) or 1 µg/mL of Anti-Human COX IV Purified (right) followed by F (ab')2 Anti-Mouse IgG eFluor® 570.Nuclei are stained with DAPI.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of COX4 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with COX4 Monoclonal Antibody (IB52C31H10), eBioscience™ (Product # 14-9775-82) at a concentration of 1 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.