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Validated using a knockout cell line

Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)

Submit a review Q&A (2)References (6)
Immunocytochemistry/ Immunofluorescence - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
  • Flow Cytometry - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
  • Immunocytochemistry/ Immunofluorescence - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)

Key features and details

  • Mouse monoclonal [9E2BD2] to SOD2/MnSOD
  • Suitable for: ICC/IF, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-SOD2/MnSOD antibody [9E2BD2]
    See all SOD2/MnSOD primary antibodies

  • Description

    Mouse monoclonal [9E2BD2] to SOD2/MnSOD

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Full length native protein (purified). This information is considered to be commercially sensitive.

  • Positive control

    • Human HDFn and HL-60 cells and Human cerebellum. ICC-IF: HAP1 cells (HAP1-SOD2 knockout cells used as negative cell line)

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Product was previously marketed under the MitoSciences sub-brand.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab110300 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
ICC/IF
Use a concentration of 1 - 5 µg/ml.
IHC-P
1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. use 1mmol EDTA pH8
Flow Cyt
Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Notes
ICC/IF
Use a concentration of 1 - 5 µg/ml.
IHC-P
1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. use 1mmol EDTA pH8
Flow Cyt
Use a concentration of 1 µg/ml.

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

Target

  • Function

    Destroys superoxide anion radicals which are normally produced within the cells and which are toxic to biological systems.

  • Involvement in disease

    Genetic variation in SOD2 is associated with susceptibility to microvascular complications of diabetes type 6 (MVCD6) [MIM:612634]. These are pathological conditions that develop in numerous tissues and organs as a consequence of diabetes mellitus. They include diabetic retinopathy, diabetic nephropathy leading to end-stage renal disease, and diabetic neuropathy. Diabetic retinopathy remains the major cause of new-onset blindness among diabetic adults. It is characterized by vascular permeability and increased tissue ischemia and angiogenesis.

  • Sequence similarities

    Belongs to the iron/manganese superoxide dismutase family.

  • Post-translational
    modifications

    Nitrated under oxidative stress. Nitration coupled with oxidation inhibits the catalytic activity.

  • Cellular localization

    Mitochondrion matrix.
  • Information by UniProt

  • Database links

    • Alternative names

      • Indophenoloxidase B antibody
      • IPO B antibody
      • IPOB antibody
      • Manganese containing superoxide dismutase antibody
      • Manganese SOD antibody
      • Manganese superoxide dismutase antibody
      • Mangano superoxide dismutase antibody
      • Mn SOD antibody
      • Mn superoxide dismutase antibody
      • MNSOD antibody
      • MVCD6 antibody
      • SOD 2 antibody
      • SOD2 antibody
      • SODM_HUMAN antibody
      • Superoxide dismutase [Mn] mitochondrial antibody
      • Superoxide dismutase [Mn], mitochondrial antibody
      • Superoxide dismutase 2 mitochondrial antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
      Immunocytochemistry/ Immunofluorescence - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)

      ab110300 staining SOD2 in wild-type HAP1 cells (top panel) and SOD2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab110300 at 5ug/ml and ab6046 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in pseudo color red).  Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
      Immunohistochemistry of Superoxide Dismutase 2 in Human cerebellum visualized with ab110300. Superoxide Dismutase 2 immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells. Note the distinctive subcellular localization of Superoxide Dismutase 2 immunoreactivity in the Purkinje cell bodies.
    • Flow Cytometry - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
      Flow Cytometry - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
      HL-60 cells were stained with 1 µg/mL ab110300 in blue or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
    • Immunocytochemistry/ Immunofluorescence - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
      Immunocytochemistry/ Immunofluorescence - Anti-SOD2/MnSOD antibody [9E2BD2] (ab110300)
      Immunocytochemistry image of Superoxide Dismutase 2 stained Human HDFn cells. The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15min). The cells were incubated with ab110300 at 5 µg/ml for 2h at room temperature or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps. DAPI was used to stain the cell nuclei (blue). Target protein locates mainly in mitochondria.

    References (6)

    Publishing research using ab110300? Please let us know so that we can cite the reference in this datasheet.

    ab110300 has been referenced in 6 publications.

    • Shang L  et al. SS-31 Protects Liver from Ischemia-Reperfusion Injury via Modulating Macrophage Polarization. Oxid Med Cell Longev 2021:6662156 (2021). PubMed: 33986918
    • Wang J  et al. Thioredoxin relieves lipopolysaccharide-induced acute kidney injury in mice by reducing inflammation, oxidative stress and apoptosis. Exp Ther Med 21:629 (2021). PubMed: 33936285
    • Sun X  et al. Neutralization of Oxidized Phospholipids Ameliorates Non-alcoholic Steatohepatitis. Cell Metab 31:189-206.e8 (2020). PubMed: 31761566
    • Abuaita BH  et al. Mitochondria-Derived Vesicles Deliver Antimicrobial Reactive Oxygen Species to Control Phagosome-Localized Staphylococcus aureus. Cell Host Microbe 24:625-636.e5 (2018). PubMed: 30449314
    • González-Fernández C  et al. Wnt Signaling Alteration in the Spinal Cord of Amyotrophic Lateral Sclerosis Transgenic Mice: Special Focus on Frizzled-5 Cellular Expression Pattern. PLoS One 11:e0155867 (2016). IHC . PubMed: 27192435
    • Zhang C  et al. Heteroplasmic mutations of the mitochondrial genome cause paradoxical effects on mitochondrial functions. FASEB J : (2012). WB ; Human . PubMed: 22925728

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Question

    IP with mouse mitochondria
    WB detects band
    IP with recombinant protein works (5 ug Ab for 700 ng protein)
    for lysate: no band, no pull-down
    Ab: lysate ratio tried are: 5 ul/50 ul (1/10) and 2ul/50 ul (1/25)
    which specific IP protocol was used? which lysis buffer was used? any additional details?
    suggest other SOD2 Abs that would work in IP

    Read More
    Answer

    Thank you for contacting us.

    The lab sent me the following information:

    The customer should use this protocol: http://www.mitosciences.com/PDF/ip.pdf
    As in this protocol we would recommend conjugating the antibody to the protein G beads prior to IP, but can also be done by adding beads to lysates/antibody mix. The detergent lauryl maltoside is ab109857 (https://www.abcam.com/10-Lauryl-Maltoside-Solution-ab109857.html)
    We would recommend 1-5 mg lysates material from cell/tissue/mitochondria prepared using the detergent specific in the protocol (lauryl maltoside).



    I hope this information is helpful to you. Please let me know if this protocol helps to get the IP to work with the endogenous samples.

    Also, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    Hi Heather, thanks a lot for your response last month to my inquiry regarding using antiSOD2 clone 9E2BD2/ab110300 in a flow cytometry assay. I obtained the antibody and attempted to stain murine cells with it, but was unable to detect any staining by flow.  In contrast, the flow plot in the product literature at  https://www.abcam.com/Superoxide-Dismutase-2-antibody-9E2BD2-ab110300.html#Superoxide-Dismutase-2-Primary-antibodies-ab110300-5.jpg looks beautiful.  So I write to ask for some additional details on how this data was generated.  In particular, any information about the following staining conditions would be really helpful: what (if any?) fixation/permeabilization procedure or reagent was used? Exactly which secondary antibody was used, and at what concentration? thanks in advance for your continued assistance.

    Read More
    Answer

    Thank you for contacting us. The cells were fixed in PFA 4% for 15 minutes. For permeabilization 90% methanol was used at -20 degrees Celcius for 30 minutes. Alternatively 0.1% Triton-X may be used and optimized. Alexa 488 was used as the seconday antibody. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

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