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Catalog Number | orb548371 |
---|---|
Category | Antibodies |
Description | COX IV COX4I1 Antibody (monoclonal, 4G11) |
Species/Host | Mouse |
Clonality | Monoclonal |
Clone Number | 4G11 |
Tested applications | FC, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Mouse IgG2b |
Immunogen | E. coli-derived human COX IV recombinant protein (Position: Q59-K169). |
Concentration | 0 |
Dilution range | Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Flow Cytometry, 1-3μg/1x106 cells |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 17 kDa |
UniProt ID | P13073 |
Storage | Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles. |
Alternative names | Cytochrome c oxidase subunit 4 isoform 1, mitochon Read more... |
Note | For research use only |
Application notes | Add 0.2ml of distilled water will yield a concentration of 500μg/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of COX IV using anti-COX IV antibody (orb548371). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human HEPG2 whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human HEK293 whole cell lysates, Lane 4: human T47D whole cell lysates, Lane 5: human CACO-2 whole cell lysates, Lane 6: human K562 whole cell lysates, Lane 7: human HeLa whole cell lysates, Lane 8: rat brain tissue lysates, Lane 9: mouse brain tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-COX IV antigen affinity purified monoclonal antibody (Catalog # orb548371) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for COX IV at approximately 17KD. The expected band size for COX IV is at 17KD.
IHC analysis of COX IV using anti-COX IV antibody (orb548371). COX IV was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COX IV Antibody (orb548371) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of COX IV using anti-COX IV antibody (orb548371). COX IV was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COX IV Antibody (orb548371) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of COX IV using anti-COX IV antibody (orb548371). COX IV was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-COX IV Antibody (orb548371) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
Flow Cytometry analysis of A431 cells using anti-COX IV antibody (orb548371). Overlay histogram showing A431 cells stained with orb548371 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-COX IV Antibody (orb548371, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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