Overview

  • Product name
    Anti-Mitofusin 2 + Mitofusin 1 antibody [3C9]
  • Description
    Mouse monoclonal [3C9] to Mitofusin 2 + Mitofusin 1
  • Host species
    Mouse
  • Specificity

    The immunogen used for this product shares 63% homology with MFN2.

    ab57602 binds to Mitofusin 1 (Mfn1) and mitofusin 2 (Mfn2) in western blot.

  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Cynomolgus monkey
  • Immunogen

    Recombinant full length protein corresponding to Human Mitofusin 1 aa 1-741.
    Database link: Q8IWA4

  • General notes

    This product was changed from ascites to tissue culture supernatant on 05 Feb 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab57602 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 Use at an assay dependent concentration. Predicted molecular weight: 84 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.

Target

Images

  • All lanes : Anti-Mitofusin 2 + Mitofusin 1 antibody [3C9] (ab57602) at 1/1000 dilution

    Lane 1 : Recombinant Human Mitofusin 1 protein (ab132635)
    Lane 2 : MFN2 OE lysate (DDK tag) 10ng

    Secondary
    All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution

    Predicted band size: 84 kDa



    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab57602 overnight at 4°C. Antibody binding was detected using the Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 at a 1/10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

    The difference in MW is due to lane one having a GST tag and lane 2 a DDK tag.

  • ICC/IF image of ab57602 stained HepG2 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 (ab57602, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

    This image was generated using a version of the antibody produced in ascites. 

  • All lanes : Anti-Mitofusin 2 + Mitofusin 1 antibody [3C9] (ab57602) at 1/1000 dilution

    All lanes : Mouse cardiomyocytes whole cell lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-mouse IgG at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 84 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocked with 5% milk for 1 hour at 25°C.

    Incubated with the primary antibody at 4°C for 13 hours in 1X TBS.

    This image was generated using a version of the antibody produced in ascites. 

    See Abreview

  • Mitofusin 2 + Mitofusin 1 antibody (ab57602) at 1ug/lane + HeLa cell lysate at 25 μg/lane.

    This image was generated using a version of the antibody produced in ascites. 

  • IHC image of ab57602 staining in human normal kidney 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 ab57602, 1µg/ml, 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.

    This image was generated using a version of the antibody produced in ascites. 

  • Overlay histogram showing HEK293 cells stained with ab57602 (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 (ab57602, 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 IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This image was generated using a version of the antibody produced in ascites. 

  • Mitofusin 2 + Mitofusin 1 was immunoprecipitated using 0.5mg Hela whole cell extract, 10ug of Mouse monoclonal to Mitofusin 1 and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10 min under agitation. No antibody was added to the control lane 2 and no extract or antibody was added to contol lane 3. Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab57602.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Bands: 84kDa:Mitofusin 1; 60kDa bead background: non specific - 48kDa: We are unsure as to the identity of this extra band.

    This image was generated using a version of the antibody produced in ascites. 

References

This product has been referenced in:
  • Hou X  et al. Mitofusin1 in oocyte is essential for female fertility. Redox Biol 21:101110 (2019). Read more (PubMed: 30690319) »
  • Yen JH  et al. Activation of dynamin-related protein 1 - dependent mitochondria fragmentation and suppression of osteosarcoma by cryptotanshinone. J Exp Clin Cancer Res 38:42 (2019). Read more (PubMed: 30691497) »
See all 89 Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
Permeabilization
Yes - 0.05% Tween 20
Specification
Pancreas
Blocking step
sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Formaldehyde

Mr. David Ivancic

Verified customer

Submitted Jul 03 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293)
Permeabilization
Yes - 0.1% Triton X100
Specification
HEK293
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 10 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Prostate cancer cell line)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Prostate cancer cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 04 2016

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell)
Specification
CHO cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 18 2015

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (10% PAGE)
Sample
Mouse Cell lysate - whole cell (Cardiomyocytes)
Specification
Cardiomyocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 30 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Cardiomyocytes)
Specification
Cardiomyocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 19 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (hepatocytes)
Specification
hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Dec 23 2014

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Sample
Mouse Tissue lysate - whole (Liver)
Specification
Liver
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 09 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12% Bis-Tris)
Sample
Cynomolgus Monkey Cell lysate - whole cell (Peripheral Blood Mononuclear Cell)
Specification
Peripheral Blood Mononuclear Cell
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Mr. Daniel Tyrrell

Verified customer

Submitted Oct 18 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293T)
Loading amount
20 µg
Specification
HEK 293T
Gel Running Conditions
Reduced Denaturing (10% Bis Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 01 2011

1-10 of 11 Abreviews

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