Anti-SOD2/MnSOD antibody [mAbcam 74231] (ab74231)

Overview

  • Product name
    Anti-SOD2/MnSOD antibody [mAbcam 74231]
    See all SOD2/MnSOD primary antibodies
  • Description
    Mouse monoclonal [mAbcam 74231] to SOD2/MnSOD
  • Tested applications
    Suitable for: Flow Cyt, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Rabbit, Horse, Guinea pig, Cow, Pig, Chimpanzee, Zebrafish, Cynomolgus monkey, Macaque monkey, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human SOD2/MnSOD aa 50-150 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab94556)

  • Positive control
    • WB: human liver; human brain; human heart; rat brain tissue lysates.
  • General notes

    This antibody clone is manufactured by Abcam.

    This monoclonal antibody to SOD2 has been knockout validated in Western blot. The expected band for SOD2 was observed in wild type cells and the band was not seen in knockout cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab74231 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
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 25 kDa).

We recommend using 3% milk as the blocking agent in Western Blot.

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

Anti-SOD2/MnSOD antibody [mAbcam 74231] images



  • Predicted band size : 25 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: SOD2 knockout HAP1 cell lysate (20 µg)
    Lane 3: Human heart cell lysate (20 µg)
    Lane 4: Rat heart cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab74231 observed at 29 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab74231 was shown to specifically react with SOD2. Wild-type and SOD2 knockout samples were subjected to SDS-PAGE. ab74231 and ab181602 (loading control to GAPDH) were diluted 1 μg/mL and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-SOD2/MnSOD antibody [mAbcam 74231] (ab74231) at 5 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Human heart tissue lysate - total protein (ab29431)
    Lane 3 : Human brain tissue lysate - total protein (ab29466)
    Lane 4 : Brain (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 25 kDa
    Observed band size : 23 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

    The Superoxide dismutase 2 protein has a predicted molecular weight of 25-kDa. However, the sequence contains a 24 residue transit peptide at the N-terminus and, when cleaved, the protein has an expected molecular weight of approximately 23-kDa.

     

    We recommend using 3% milk as the blocking agent in Western Blot.

  • Overlay histogram showing A549 cells stained with ab74231 (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 (ab74231, 1µg/1x106 cells) 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 A549 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 25 kDa


    Exposure time : 12 minutes

References for Anti-SOD2/MnSOD antibody [mAbcam 74231] (ab74231)

ab74231 has not yet been referenced specifically in any publications.

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