Overview

  • Product name

    Anti-Mitochondria antibody [113-1]
    See all Mitochondria primary antibodies
  • Description

    Mouse monoclonal [113-1] to Mitochondria
  • Host species

    Mouse
  • Tested applications

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

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Human cell homogenate

  • Positive control

    • WB: HeLa cell lysate. ICC/IF: HeLa and U87 cells. IHC-P: Human breast cancer tissue. FC: HepG2 cells.
  • General notes


    This antibody is an excellent marker for Human cells in xenographic model research.

Properties

Applications

Our Abpromise guarantee covers the use of ab92824 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
WB 1/1000. Detects a band of approximately 60 kDa.
IHC-P 1/1000.
ICC/IF 1/800.
Flow Cyt 1/100.

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

Target

  • Relevance

    Mitochondria 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 localization

    Mitochondrial

Images

  • Immunostained human cells in spleen and temporal artery (TA) xenograft sections demonstrate persistent human PBMC colonization at 28 days

    Mitochondria in spleen (B) and temporal artery xenografts (E) were detected using ab92824 at 1/400 dilution in immunohistochemical analysis.

    Magnification: 400X for spleen, 200X for TA grafts and 800X for insets.

    (From Figure 2 of Chen et al)

  • Anti-Mitochondria antibody [113-1] (ab92824) at 1/1000 dilution + HeLa lysate

    Observed band size: 60 kDa
    why is the actual band size different from the predicted?

  • ICC/IF image of ab92824 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 (ab92824, 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.

  • IHC image of ab92824 staining in Breast Cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab92824, 1/1000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Overlay histogram showing HepG2 cells stained with ab92824 (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 (ab92824, 1/100 dilution) 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 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 HepG2 cells fixed with 4% paraformaldehyde (10 min) permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ab92824 staining mitochondria in the Human U87 glioblastoma cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 10% NBF, permeabilized with Triton X-100 and blocked with 2% serum for 90 minutes at 22°C. Samples were incubated with primary antibody (1/1000 in 0.2% BSA + 2% NGS) for 15 hours at 4°C. An Alexa Fluor® 568-conjugated Goat anti-mouse IgG1 polyclonal was used as the secondary antibody (1/500). Co stained with ActinGreen, 2 drops/ml PBS, 10 min and Hoechst 2ug/ml in H2O 10 min

    See Abreview

References

This product has been referenced in:

  • Simeone K  et al. Paraffin-embedding lithography and micro-dissected tissue micro-arrays: tools for biological and pharmacological analysis of ex vivo solid tumors. Lab Chip 19:693-705 (2019). Read more (PubMed: 30671574) »
  • Baklaushev VP  et al. Tissue Engineered Neural Constructs Composed of Neural Precursor Cells, Recombinant Spidroin and PRP for Neural Tissue Regeneration. Sci Rep 9:3161 (2019). Read more (PubMed: 30816182) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 16 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Human Cultured Cells (Insulinoma cell)
Permeabilization
Yes - 0.25% Triton-X
Specification
Insulinoma cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 26°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 03 2019

This product is known to not work in this application or species.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Mouse Brain with Human Tumor Cells)
Specification
Mouse Brain with Human Tumor Cells
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Aug 09 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Human cells in rat heart xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6
Specification
Human cells in rat heart xenograft
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Srishti Bhutani

Verified customer

Submitted Feb 02 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
skin tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 05 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rabbit Cell (RAB-9 skin Fibroblast)
Permeabilization
Yes - 0.25% Triton X-100
Specification
RAB-9 skin Fibroblast
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Paraformaldehyde

Miss. Michaela Carroll

Verified customer

Submitted Feb 19 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U87 glioblastoma)
Permeabilization
Yes - triton x 100
Specification
U87 glioblastoma
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
10% NBF, 10 min

Virginia Hoglund

Verified customer

Submitted Jul 27 2015

Answer

Thank you for contacting us.

Unfortunately we will not be able to provide this antibody conjugated with Biotin or any other conjugate. Could you however let us know if we can provide further help in your search e.g. if you are trying to look for mitochondrial staining then any antibody against mitochondrial marker would be helpful. This will actually broadens the search results.
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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Answer

I have a further update relating to your enquiry.

The Anti-Mitochondria antibody [113-1], ab92824, should also be suitable for the customers experiments. It has been shown not to cross react with either mouse or rat. Characterization in both Western blots and IHC has been performed in human to yield positive results and rat and mouse to yield negative results. Furthermore, the antibody has been used in xenograft mouse models to specifically stain only transplanted human cells.

Please just let me know should you need any further information.

Read More

Answer

Thank you for contacting us and your interest in our products.

As discussed over the phone, the anti-actin antibody ab3280 has been shown to reacts with all six known vertebrate isoforms of actin. It also reacts with two cytoplasmic actins (beta, lambda) which are highly homologous to one another but which differ from the muscle actins at about 25 amino acid residues.

I have also checked whether the exact protein detected by the anti-Mitochondria antibody (ab92824) has been established. Unfortunately we do not exactly know which proteins the antibody is detecting as a human mitochondrial extract was used as antigen and we therefore suspect that the target is not a single protein, but rather likely multiple proteins that are specific to mitochondria. Sorry we can't be of further help in this instance.

I was interested to hear you have been using these antibodies with good results in EM. We and our customers would be very interested if you would share these results. We have an Abreview system where if you share your results with any of our products you will receive credit to use against a future Abcam order or Amazon vouchers. More information on this system can be found from the following link:

www.abcam.com/abreviews

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to ask.

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Question
Answer

Thank you for contacting us.

I apologize for the delay; the concentration of lot GR29268-9 is 1 mg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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1-10 of 16 Abreviews or Q&A

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