Overview

  • Product name

    Anti-Mitofilin antibody [EPR8749]
    See all Mitofilin primary antibodies
  • Description

    Rabbit monoclonal [EPR8749] to Mitofilin
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    corresponding to Human Mitofilin aa 50-150.

  • Positive control

    • WB: HAP1, HepG2, HeLA whole cel lysates; human heart, fetal heart and skeletal muscle tissues IHC-P: Human thyroid cancer, hepatocellular cancer, colon and testis tissue ICC/IF: HeLa cells FC: HeLa cells
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab137057 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 - 1/10000. Detects a band of approximately 80-84 kDa (predicted molecular weight: 84 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/100 - 1/500.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Cellular localization

    Mitochondrion inner membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cell proliferation inducing protein 52 antibody
    • Cell proliferation-inducing gene 4/52 protein antibody
    • Heart muscle protein antibody
    • HMP antibody
    • IMMT antibody
    • IMMT_HUMAN antibody
    • Inner membrane protein mitochondrial antibody
    • Inner mitochrondial membrane antibody
    • MICOS complex subunit MIC60 antibody
    • MINOS2 antibody
    • Mitochondrial inner membrane organizing system 2 antibody
    • Mitochondrial inner membrane protein antibody
    • Mitofilin antibody
    • Motor protein antibody
    • p87 antibody
    • p87/89 antibody
    • p89 antibody
    • pig4 antibody
    • PIG52 antibody
    • Proliferation-inducing gene 4 antibody
    see all

Images

  • All lanes : Anti-Mitofilin antibody [EPR8749] (ab137057) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : IMMT knockout HAP1 whole cell lysate
    Lane 3 : HepG2 whole cell lysate
    Lane 4 : Human Heart whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 84 kDa
    Observed band size: 84 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab137057 observed at 84 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab137057 was shown to specifically react with IMMT in wild-type HAP1 cells as signal was lost in IMMT knockout cells. Wild-type and IMMT knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab137057 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human thyroid cancer tissue sections labeling Mitofilin with purified ab137057 at 1/250 dilution (0.508 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • All lanes : Anti-Mitofilin antibody [EPR8749] (ab137057) at 1/10000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : Human fetal heart lysates
    Lane 3 : Human skeletal muscle lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 84 kDa
    Observed band size: 84 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular cancer tissue sections labeling Mitofilin with purified ab137057 at 1/250 dilution (0.508 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • All lanes : Anti-Mitofilin antibody [EPR8749] (ab137057) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : Human fetal heart tissue lysate
    Lane 3 : Human fetal muscle tissue lysate
    Lane 4 : A673 cell lysate
    Lane 5 : Saos 2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution

    Predicted band size: 84 kDa

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Mitofilin with purified ab137057 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Mitofilin with purified ab137057 at 1/100 dilution (1.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Mitofilin with ab137057 (unpurified) at 1/100 dilution.

  • Immunohistochemical analysis of paraffin embedded Human testis tissue labelling Mitofilin with ab137057 (unpurified) at 1/100 dilution.

  • Immunofluorescent staining of HeLa cells labelling Mitofilin with ab137057 (unpurified) at 1/250 dilution.

References

This product has been referenced in:

  • Matsuhashi T  et al. Mitochonic Acid 5 (MA-5) Facilitates ATP Synthase Oligomerization and Cell Survival in Various Mitochondrial Diseases. EBioMedicine 20:27-38 (2017). WB ; Cow . Read more (PubMed: 28579242) »
  • Sastri M  et al. Sub-mitochondrial localization of the genetic-tagged mitochondrial intermembrane space-bridging components Mic19, Mic60 and Sam50. J Cell Sci 130:3248-3260 (2017). Read more (PubMed: 28808085) »
See all 4 Publications for this product

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