Recombinant
RabMAb

Recombinant Anti-SIRT3 antibody [EPR19755] (ab217319)

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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

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

    This product is a recombinant rabbit monoclonal antibody.

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.

    References

    This product has been referenced in:

    • 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). Read more (PubMed: 31247208) »
    • 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). Read more (PubMed: 30487699) »
    See all 2 Publications for this product

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

    Customer reviews and Q&As

    Show All Reviews Q&A
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    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

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    Verified customer

    Submitted Apr 23 2019

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