Overview

  • Product nameAnti-Mitochondria antibody [MTC02]
    See all Mitochondria primary antibodies
  • Description
    Mouse monoclonal [MTC02] to Mitochondria
  • Tested applicationsIHC-FrFl, ICC/IF, ICC, IHC-P, IHC-Fr, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human, Pig, Rhesus monkey, African Green Monkey
  • Immunogen

    Semi purified mitochondrial preparation.

  • EpitopeThis antibody recognizes a 60 kDa non glycosylated protein component of mitochondria found in human cells.
  • Positive control
    • HeLa cells. Liver tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab3298 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
IHC-FrFl Use at an assay dependent concentration.
ICC/IF Use at an assay dependent dilution. Used at a dilution of 1/50 for 2 hours (see Abreview for further information).
ICC Use at an assay dependent dilution.
IHC-P Use a concentration of 1 - 2 µg/ml. PubMed: 18167556Samples require boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10 - 20 min followed by cooling at RT for 20 min. This antibody can be used to stain themitochondria in cell / tissue preparations and can be used as a marker of the mitochondria in subcellular fractions.
IHC-Fr Use at an assay dependent dilution.
WB Use a concentration of 1 - 2 µg/ml. This antibody detects a band of 60 kDa, which corresponds to the predicted molecular weight of the non glycosylated protein component of mitochondria.
Flow Cyt Use 1µg for 106 cells.

Target

  • RelevanceMitochondria are the power house of the cell. They are distinct organelles with two membranes. Usually they are rod shaped, however they can be round. The outer membrane limits the organelle and the inner membrane is thrown into folds or shelves that project inward and are called "cristae mitochondriales".
  • Cellular localizationMitochondrial

Anti-Mitochondria antibody [MTC02] images

  • Overlay histogram showing HEK293 cells stained with ab3298 (red line). The cells were fixed with 100% 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 (ab3298, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Mitochondria were detected by Immunofluorescence in U373 human glioblastoma cells using Mitochondrial Marker antibody (ab3298).

  • ab3298 at 0.2µg/ml staining Mitochondria-Mitochondrial Marker from human cardiac muscle sections by Immunohistochemistry (Paraffin-embedded sections). The antibody was incubated with the tissue for 2 hours and then detected with an Alexa-Fluor® 488 conjugated anti-mouse IgG secondary antibody.

    This image is courtesy of an Abreview submitted by an anonymous researcher on 12 December 2005.

    See Abreview

  • ab3298 staining HEK 293 cells by ICC/IF.  Following paraformaldehyde fixation and permeabilisation with 0.5%TX100 the cells were blocked with 5% serum (for 20 hours at 25°C).  The primary antibody was diluted 1/100 in PBS, 2% goat serum, 0.1% TX100 and incubated for 1 hour at 25°C.  An Alexa Fluor® 546 conjugated goat anti-mouse was used as the secondary.

    Mitochondria are labeled with red fluorescence, and the nuclei were counterstained with DAPI.

    See Abreview

  • ab3298 at 1/200 dilution staining mitochondria in human foreskin fibroblasts by immunocytochemistry/ immunofluorescence. Sections were paraformaldehyde fixed, permeabilized in 0.1% saponin prior to blocking in BSA (1%) + normal goat serum (2%) for 30 minutes at 20°C and then incubated with ab3298 for 1 hour at 20°C. Alexa fluor® 594 goat polyclonal to mouse Ig, diluted 1/1000, was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab3298 stained HeLa cells. The cells were 4% PFA 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 (ab3298, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) 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.

  • ab3298 staining mitochondria in Mongolian gerbil brain tissue by Immunohistochemistry - Free Floating.
    ab3298 used at a 1/300 dilution for 16 hours at 4°C with 1% BSA, 0.3% Triton X-100, 0.1% Saponin. The secondary used was a goat anti-mouse IgG1 DyLight 649 used at a 1/300 dilution.

    See Abreview

References for Anti-Mitochondria antibody [MTC02] (ab3298)

This product has been referenced in:
  • Taha MS  et al. Subcellular fractionation and localization studies reveal a direct interaction of the fragile X mental retardation protein (FMRP) with nucleolin. PLoS One 9:e91465 (2014). ICC/IF ; Human . Read more (PubMed: 24658146) »
  • Hall CJ  et al. Epidermal cells help coordinate leukocyte migration during inflammation through fatty acid-fuelled matrix metalloproteinase production. Nat Commun 5:3880 (2014). Zebrafish . Read more (PubMed: 24852213) »

See all 22 Publications for this product

Product Wall


According to the datasheet, this antibody is a purified antibody, not an ascites antibody, with a concentration of 500 µl at 1 mg/ml.
Moreover, I could not finds any hints that we have changed the formulation from ascites to purified recently...

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Although the anti-mitochondria antibody ab3298 has been used in many published studies, including the ones listed in the references section of the datasheet, the identity of the protein that the antibody recognizes is not known. None of the references ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Glioblastoma)
Specification Glioblastoma
Fixative Paraformaldehyde
Permeabilization Yes - Triton X-100 1%
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 13 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human MDM)
Specification human MDM
Fixative Paraformaldehyde
Permeabilization Yes - TRITON-X (0.1 %)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 05 2013

This antibody is indeed guaranteed to work in IHC and should label the mitochondria in cells. However, we have not used this antibody to quantify mitochondria in cells before and therefore cannot guarantee that this will work. We do not have much exper...

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Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guar...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rhesus monkey Cell (FRhK-4 cells)
Specification FRhK-4 cells
Fixative Paraformaldehyde
Permeabilization Yes - 0.2% Triton-X100 in PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Username

Abcam user community

Verified customer

Submitted Nov 07 2012

Thank you once again for your enquiries and for confirming the product code as

I am sorry to confirm this antibody is sourced externally. I have requested information regarding which mitochondrial protein it detects, and they regrettably hav...

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Yes, the antibody would be guaranteed.

If you should have any problems, please report itto us within 6 month of the purchase, and we would offer either a replacement, credit or refund.

I wish you good luck!

Thank you for your enquiry. That customer review is the only mouse data we have for immunohistochemistry or immunocytochemistry. We do have flow cytometry data for a PFA-fixed human cell line, and we expect PFA-fixed mouse cells or PFA-fixed mouse tiss...

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