MitoSciences (MS101c)

Anti-Complex I Immunocapture antibody [18G12BC2] (ab109798)

Overview

  • Product nameAnti-Complex I Immunocapture antibody [18G12BC2]
  • Description
    Mouse monoclonal [18G12BC2] to Complex I Immunocapture
  • Tested applicationsIP, ICC, In-Cell ELISA, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Tissue, cells or virus corresponding to Complex I Immunocapture. Purified cow mitochondrial complex I.

  • Positive control
    • fibroblasts, HL-60 cells, tissue mitochondria

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C. Do not freeze.
  • Storage bufferPreservative: 0.02% Sodium azide
    Constituent: 99% HBS
  • Concentration information loading...
  • PurityIgG fraction
  • Purification notesNear homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
  • ClonalityMonoclonal
  • Clone number18G12BC2
  • IsotypeIgG2b
  • Light chain typekappa
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab109798 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
IP Use at 100-1 µg/mg of lysate. 100 µg mAb can capture at least 25 µg complex I from 1 mg solubilized bovine heart mitochondria.
ICC Use at an assay dependent dilution.
In-Cell ELISA Use a concentration of 1 µg/ml.
Flow Cyt Use 2µg for 106 cells.

Target

  • RelevanceComplex I, or NADH ubiquinone oxidoreductase, is a large protein complex of 950,000 Da molecular weight made up by 45 to 46 different subunits. A total of seven of the subunits of the complex are encoded by mitochondrial DNA, while the remainder subunits are nuclear encoded, which are translated in the cytosol and translocated into the organelle for assembly at the inner membrane. The enzyme complex catalyses electron entry from NADH via a flavin (FMN) and several non-heme iron centers. Complex I is sensitive to a wide range of inhibitors, many of which are pesticides or other common environmental toxins, such as rotenone. Complex I dysfunction is a common cause of genetic OXPHOS defects. Altered functioning of this complex is also thought to contribute to several neurological disorders including Parkinson’s disease and schizophrenia. Also, there is evidence of Complex I involvement in diabetes.

Anti-Complex I Immunocapture antibody [18G12BC2] images

  • Immunocytochemistry image of ab109798 stained fibroblasts cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with the antibody (ab109798, 1 µg/mL) for 2 hours at room temperature or over night at 4°C. The secondary antibody was (green) Alexa Fluor® 4884 goat anti-mouse IgG (H+L) at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. The target protein locates to the mitochondria.
  • HL-60 cells were stained with 1 µg/mL Complex I antibody ab109798 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
  • Overlay histogram showing HepG2 cells stained with ab86750 (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. The cells were then incubated with the antibody (ab867508, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Complex I Immunocapture antibody [18G12BC2] (ab109798)

This product has been referenced in:
  • Zhang J  et al. Extended wakefulness: compromised metabolics in and degeneration of locus ceruleus neurons. J Neurosci 34:4418-31 (2014). Mouse . Read more (PubMed: 24647961) »
  • Willis JH  et al. Isolated deficiencies of OXPHOS complexes I and IV are identified accurately and quickly by simple enzyme activity immunocapture assays. Biochim Biophys Acta 1787:533-8 (2009). Read more (PubMed: 19041632) »

See all 10 Publications for this product

Product Wall

The image is not of a blot but of a gel stained for total protein. The annotations refer to subunits of mouse brain mitochondria Complex I, identified by mass spec as described in the reference at this link: http://www.mcponline.org/content/4/1/84.full...

Read More

This antibody is an immunocapture antibody which can pull down the whole complex I from a given sample. It works also as a ICC and Flow antibody. It's a weak WB antibody, light up a 8kda subunit of complex I but we have difficulty to identify which sub...

Read More

whole (45 subunit, 900 kDa) active bovine Complex I - this was shown with mass spec.

It is excellent at binding native, active complex I but has proven difficult to identify the exact subunit to which it binds.

Thank you for contacting Abcam.

I have attached a paper that looks at the structure of complex 1 and Table 1 has a complete list of the subunits and I believe (but I am no expert in this field) that S1 is also known as NDUFS1.

Pleas...

Read More

Thank you very much for your phone call.
I have had discussion with my colleague in lab. They have suggested that the antibody was tested only with lysates and cells as presented on the datasheet. No other tested data is available for this product...

Read More

Thank you for contacting us. I am not sure which publication you are looking at however I have the following publication from Sashi Nadanaciva; http://www.mitosciences.com/PDF/MitoSciences_OXPHOS_Screen_Paper.pdf In this article MS101c is u...

Read More

Thank you for contacting us. I suppose you have a question about ab109798; Anti-Complex I Immunocapture antibody. This antibody detects the presence of Complex I in cell sample; this unfortunately is not suitable for assaying the enzymatic activity...

Read More

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.mi...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"