RecombinantRabMAb

Recombinant Anti-SIRT3 antibody [EPR19755] (ab217319)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19755] to SIRT3
  • Suitable for: WB, IHC-P
  • Reacts with: Human

Overview

  • Product name

    Anti-SIRT3 antibody [EPR19755]
    See all SIRT3 primary antibodies

  • Description

    Rabbit monoclonal [EPR19755] to SIRT3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human SIRT3 aa 300 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9NTG7

  • Positive control

    • WB: HEK-293, HeLa, HepG2 and HT-29 whole cell lysates; Human fetal brain, fetal heart, fetal kidney and fetal liver lysates. IHC-P: Human liver, hepatocellular carcinoma, colon and colon adenocarcinoma tissues.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab217319 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. Detects a band of approximately 28 kDa (predicted molecular weight: 43 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    NAD-dependent protein deacetylase. Activates or deactivates mitochondrial target proteins by deacetylating key lysine residues. Known targets include ACSS1, IDH, GDH, SOD2, PDHA1, LCAD, SDHA and the ATP synthase subunit ATP5O. Contributes to the regulation of the cellular energy metabolism. Important for regulating tissue-specific ATP levels.

  • Tissue specificity

    Widely expressed.

  • Sequence similarities

    Belongs to the sirtuin family. Class I subfamily.
    Contains 1 deacetylase sirtuin-type domain.

  • Post-translational
    modifications

    Processed by mitochondrial processing peptidase (MPP) to give a 28 kDa product. Such processing is probably essential for its enzymatic activity.

  • Cellular localization

    Mitochondrion matrix.
  • Information by UniProt

  • Database links

    • Alternative names

      • hSIRT 3 antibody
      • hSIRT3 antibody
      • Mitochondrial nicotinamide adenine dinucleotide dependent deacetylase antibody
      • NAD dependent deacetylase sirtuin 3 mitochondrial antibody
      • NAD-dependent protein deacetylase sirtuin-3, mitochondrial antibody
      • Regulatory protein SIR2 homolog 3 antibody
      • Silent mating type information regulation 2 S.cerevisiae homolog 3 antibody
      • Sir 2 like 3 antibody
      • SIR 2 like protein 3 antibody
      • SIR 3 antibody
      • SIR2 L3 antibody
      • Sir2 like 3 antibody
      • SIR2 like protein 3 antibody
      • SIR2-like protein 3 antibody
      • SIR2L3 antibody
      • SIR3_HUMAN antibody
      • SIRT 3 antibody
      • SIRT3 antibody
      • Sirtuin 3 antibody
      • Sirtuin silent mating type information regulation 2 homolog 3 (S. cerevisiae) antibody
      • Sirtuin type 3 antibody
      • Sirtuin3 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)

      Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling SIRT3 with ab217319 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on human hepatocellular carcinoma is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      According to the literature [PMID: 24774224]: IHC and WB studies both showed a decreased expression of SIRT3 in tumor tissues compared with peritumor tissues. Our IHC-P results also showed decreased expression on about 58% of hepatocellular carcinoma.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)

      Immunohistochemical analysis of paraffin-embedded human liver tissue labeling SIRT3 with ab217319 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on human liver is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)

      Immunohistochemical analysis of paraffin-embedded human colon tissue labeling SIRT3 with ab217319 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on human colon is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)

      Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma tissue labeling SIRT3 with ab217319 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on human well-differentiated colon adenocarcinoma is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Our IHC-P results show that about 75% of well-differentiated colon carcinoma show increased expression.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT3 antibody [EPR19755] (ab217319)

      Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma tissue labeling SIRT3 with ab217319 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human poor- differentiated colon adenocarcinoma is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      According to the literature [PMID: 25105144]: SIRT3 expression level was found to be significantly associated with the lymph node metastasis and tumor stages. Our IHC-P results show that about 25% of poor-differentiated colon carcinoma show decreased expression.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-SIRT3 antibody [EPR19755] (ab217319)
      Western blot - Anti-SIRT3 antibody [EPR19755] (ab217319)
      All lanes : Anti-SIRT3 antibody [EPR19755] (ab217319) at 1/1000 dilution

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 4 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 43 kDa
      Observed band size: 28 kDa
      why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The expression profile observed is consistent with what has been described in the literature (PMID:12186850).

    • Western blot - Anti-SIRT3 antibody [EPR19755] (ab217319)
      Western blot - Anti-SIRT3 antibody [EPR19755] (ab217319)
      All lanes : Anti-SIRT3 antibody [EPR19755] (ab217319) at 1/1000 dilution

      Lane 1 : Human fetal brain lysate
      Lane 2 : Human fetal heart lysate
      Lane 3 : Human fetal kidney lysate
      Lane 4 : Human fetal liver lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 43 kDa
      Observed band size: 28 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1-3: 3 minutes; Lane 4: 15 seconds.

    • Anti-SIRT3 antibody [EPR19755] (ab217319)
      Anti-SIRT3 antibody [EPR19755] (ab217319)

    References (5)

    Publishing research using ab217319? Please let us know so that we can cite the reference in this datasheet.

    ab217319 has been referenced in 5 publications.

    • He Q  et al. Associations of sirtuins with clinicopathological variables and prognosis in human ovarian cancer. Oncol Lett 19:3278-3288 (2020). PubMed: 32256823
    • Hou L  et al. ABT737 enhances ovarian cancer cells sensitivity to cisplatin through regulation of mitochondrial fission via Sirt3 activation. Life Sci 232:116561 (2019). PubMed: 31247208
    • Gao J  et al. Lithocarpus polystachyus Rehd. leaves aqueous extract protects against hydrogen peroxide-induced SH-SY5Y cells injury through activation of Sirt3 signaling pathway. Int J Mol Med 42:3485-3494 (2018). PubMed: 30320335
    • Xu WY  et al. Zinc finger E-box-binding homeobox 1 mediates aerobic glycolysis via suppression of sirtuin 3 in pancreatic cancer. World J Gastroenterol 24:4893-4905 (2018). PubMed: 30487699
    • Lu AQ  et al. Upregulation of miR-137 reverses sorafenib resistance and cancer-initiating cell phenotypes by degrading ANT2 in hepatocellular carcinoma. Oncol Rep 37:2071-2078 (2017). PubMed: 28350139

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry free floating abreview for Anti-SIRT3 antibody [EPR19755]

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Human Tissue sections (Brain (cortex))
    Specification
    Brain (cortex)
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 20 2020

    Western blot abreview for Anti-SIRT3 antibody [EPR19755]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Guinea pig Tissue lysate - other (fetal heart)
    Gel Running Conditions
    Reduced Denaturing (4-15% precast gradient gel)
    Loading amount
    20 µg
    Specification
    fetal heart
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 23 2019

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