Overview

  • Product name

    Anti-Complex IV Immunocapture antibody [31E91B82G9]
  • Description

    Mouse monoclonal [31E91B82G9] to Complex IV Immunocapture
  • Host species

    Mouse
  • Specificity

    100 µg ab109863 can capture at least 10 µg Complex IV (COX) from 1 mg solubilized Bovine heart mitochondria.
  • Tested applications

    Suitable for: ICC/IF, IP, Flow Cytmore details
    Unsuitable for: IHC-P or WB
  • Species reactivity

    Reacts with: Cow, Human
  • Immunogen

    Full length native protein (purified). This information is considered to be commercially sensitive.

  • Positive control

    • Bovine heart and Human mitochondria IF/ICC: HeLa cell line
  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab109863 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
IP Use at 100-1 µg/mg of lysate.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

  • Application notes
    Is unsuitable for IHC-P or WB.
  • Target

    • Relevance

      Complex IV, also called cytochrome c oxidoreductase or cytochrome c oxidase, is a complex of 13 different subunits, three of which (I, II and III) are encoded on mitochondrial DNA and the remainder in the nuclear DNA. The complex contains two heme groups (a and a3) and two copper atoms as prosthetic groups. Genetic alterations of this enzyme complex are a common cause of OXPHOS diseases and the enzyme is altered in patients with Alzheimer’s disease. Also, there are reports of reduced amounts of this complex in hypoxic cancer cells.
    • Cellular localization

      Mitochondrial

    Images

    • ICC/IF image of ab109863 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab109863, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Overlay histogram showing HepG2 cells stained with ab109863 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109863, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Staining of Complex IV Lane 1: 0.1 mg Human mitochondrial lysate Lane 2: 0.1 mg Bovine mitochondrial lysate Lane 3: 0.025 mg Human mitochondrial lysate Lane 4: 0.025 mg Bovine mitochondrial lysate Immunoprecipitated using ab109863 at 100 µg/mg lysate

    References

    This product has been referenced in:

    • Escobar-Alvarez S  et al. Inhibition of human peptide deformylase disrupts mitochondrial function. Mol Cell Biol 30:5099-109 (2010). Read more (PubMed: 20805355) »
    • Nadanaciva S  et al. Target identification of drug induced mitochondrial toxicity using immunocapture based OXPHOS activity assays. Toxicol In Vitro 21:902-11 (2007). Read more (PubMed: 17346924) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Answer

    Vielen Dank für Ihre Email.

    Wir haben das Lot noch nicht rezeptionniert. Ich werde mich aber sogleich bei Ihnen melden, wenn wir es in unser Lager aufgenommen haben. (Ich habe einen täglichen Reminder in meinen Kalendar getan)

    Vielen Dank für Ihre Geduld.

    Read More

    Answer

    Vielen Dank für Ihren Anruf.

    Das Labor hat mir auch bestätigt, dass es zwischen x keinen Unterschied bei der Produktion gab. In der Tat kommen beide dieser Chargen von der gleichen ursprünglichen Produktion. Ich weiss daher leider nicht, woher das Problem kommen könnte. Was denken Sie, könnte eine Ursache dieses Hintergrundes sein? Wie koppeln Sie den Antikörper auf die Beads? Der ab109863 hat ja Azide in dem Puffer, dies kann Kopplungen basierend auf Aminen beeinträchtigen.

    Das Labor hat mir auch bestätigt, dass eine neue Charge vorhanden ist. Diese Charge wäre in den nächsten Tagen erhältlich.

    Bitte lassen Sie mich wissen, wie Sie fortfahren möchten und ob Sie andere Fragen haben. Ich freue mich wieder von Ihnen zu hören.

    Read More

    Answer

    Thank you for contacting us. Please click these links to find the suitable antibodies. Type the MS code in Abcam search box to get the Abcam catalogue number. Complex I http://www.mitosciences.com/complex_i.html Complex IV http://www.mitosciences.com/complex_iv.html Complex III http://www.mitosciences.com/complex_iii.html Complex II http://www.mitosciences.com/complex_ii.html We also have Immuno capture kits available https://www.abcam.com/index.html?pageconfig=resource&rid=13917 I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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