|Applications||IHC, ICC, WB|
|Product Name||Anti-MTCO1/MT-CO1 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Cytochrome c oxidase subunit 1(MT-CO1) detection. Tested with WB, IHC-P, ICC in Human.|
|Cite This Product||Anti-MTCO1/MT-CO1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1317-1)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence at the C-terminus of human MTCO1(501-514aa PYHTFEEPVYMKS).|
Assay Dilutions Overview
Immunocytochemistry , 0.5-1μg/ml, Human, -
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Western blot, 0.1-0.5μg/ml, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P) and ICC.
Images And Assay Conditions
Figure 2. IHC analysis of MTCO1 using anti-MTCO1 antibody (PA1317-1).
MTCO1 was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1Î¼g/ml rabbit anti-MTCO1 Antibody (PA1317-1) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. IHC analysis of MTCO1 using anti-MTCO1 antibody (PA1317-1).
MTCO1 was detected in immunocytochemical section of C6 Cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1Î¼g/ml rabbit anti-MTCO1 Antibody (PA1317-1) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 1. Western blot analysis of MTCO1 using anti- MTCO1 antibody (PA1317-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The samples were loaded under reducing conditions.
Lane 1: human HeLa mitochondria lysates at 20ug,
Lane 2: human HeLa whole cell lysates at 20ug,
Lane 3: human Caco-2 whole cell lysates at 50ug,
Lane 4: rat heart tissue lysates at 50ug.
Lane 5: mouse heart tissue lysates at 50ug.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 60 minutes. Blocked the membrane with 5% Non-fat Milk in TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- MTCO1 antigen affinity purified polyclonal antibody (Catalog # PA1317-1) at 0.5 Î¼g/mL overnight at 4Â°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for MTCO1 at approximately 37KD. The expected band size for MTCO1 is at 57KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Cytochrome c oxidase subunit 1|
|Alternative Names||Cytochrome c oxidase subunit 1;188.8.131.52;Cytochrome c oxidase polypeptide I;MT-CO1;COI, COXI, MTCO1;|
|Subcellular Localization||Mitochondrion inner membrane; Multi-pass membrane protein.|
|Molecular Weight||57041 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Cytochrome c oxidase is the component of the respiratory chain that catalyzes the reduction of oxygen to water. Subunits 1- 3 form the functional core of the enzyme complex. CO I is the catalytic subunit of the enzyme. Electrons originating in cytochrome c are transferred via the copper A center of subunit 2 and heme A of subunit 1 to the bimetallic center formed by heme A3 and copper B.|
*You can search these to find other products in these research areas.
|Background||Cytochrome c oxidase subunit I(CO1 or MTCO1) is 1 of 3 mitochondrial DNA(mtDNA) encoded subunits(MTCO1, MTCO2, MTCO3) of respiratory Complex IV. Complex IV is located within the mitochondrial inner membrane and is the third and final enzyme of the electron transport chain of mitochondrial oxidative phosphorylation. It is composed of 13 polypeptides. Subunits I, II, and III(MTCO1, MTCO2, MTCO3) are encoded by mtDNA while subunits IV, Va, Vb, VIa, VIb, VIc, VIIa, VIIb, VIIc, and VIII are nuclear encoded. The cytochrome c oxidase family of enzymes have 4 redox centers, 2 hemes and 2 copper centers. In mitochondrial Complex IV, the 2 hemes are a and a3 and the 2 coppers are CuA and CuB. The 2 hemes and CuB are bound to subunit I. Acin-Perez et al.(2003) identified a cell line containing single and double missense mutations in the cytochrome c oxidase(COX) subunit I gene of mouse mitochondrial DNA. And they hypothesized that deleterious mutations can arise and become predominant; cultured cells can maintain several mtDNA haplotypes at stable frequencies; the respiratory chain has little spare COX capacity; and that the size of a cavity in the vicinity of val421 in MTCO1I of animal COX may affect the function of the enzyme.|
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Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at firstname.lastname@example.org for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact email@example.com
Q: What should I use for negative control?A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signalA: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?A: You can find the immunogen sequence under "
Q: What is the expected band size? Why is it different than the observed band size?A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?A: Some common names include but are not limited to coxi antibody, mtco1 antibody