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使用敲除细胞株进行验证

Anti-SOD2/MnSOD抗体[9E2BD2] (ab110300)

Submit a review Q&A (2)References (8)
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

概述

  • 产品名称

    Anti-SOD2/MnSOD抗体[9E2BD2]
    参阅全部 SOD2/MnSOD 一抗

  • 描述

    小鼠单克隆抗体[9E2BD2] to SOD2/MnSOD

  • 宿主

    Mouse

  • 经测试应用

    适用于: ICC/IF, IHC-P, Flow Cytmore details

  • 种属反应性

    与反应: Human

  • 免疫原

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

  • 阳性对照

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

  • 常规说明

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab110300于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
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.
说明
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.

靶标

  • 功能

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

  • 疾病相关

    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.

  • 序列相似性

    Belongs to the iron/manganese superoxide dismutase family.

  • 翻译后修饰

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

  • 细胞定位

    Mitochondrion matrix.
  • Information by UniProt

  • 数据库链接

    • 别名

      • 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

    图片

    • 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.

    文献 (8)

    发表研究结果有使用 ab110300？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab110300 被引用在 8 文献中.

    • Cheng CF  et al. Therapeutic Effects of Plant Extracts of Anoectochilus roxburghii on Side Effects of Chemotherapy in BALB/c Breast Cancer Mice. Plants (Basel) 12:N/A (2023). PubMed: 37447055
    • 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
    • Pedersen BA  et al. Novel remodeling of the mouse heart mitochondrial proteome in response to acute insulin stimulation. Nutr Metab Cardiovasc Dis 25:1152-8 (2015). PubMed: 26610654
    • Zhang C  et al. Heteroplasmic mutations of the mitochondrial genome cause paradoxical effects on mitochondrial functions. FASEB J : (2012). WB ; Human . PubMed: 22925728

    客户评价及客户问答

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    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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