Recombinant
RabMAb

Recombinant Anti-SIRT5 antibody [EPR23787-116] (ab259967)

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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 [EPR23787-116] to SIRT5
  • Suitable for: IP, WB, IHC-P, Flow Cyt
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-SIRT5 antibody [EPR23787-116]
    See all SIRT5 primary antibodies

  • Description

    Rabbit monoclonal [EPR23787-116] to SIRT5

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human SIRT5 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9NXA8

  • Positive control

    • WB: HepG2, Huh7, HeLa, HEK-293T, Hepa1-6, RAW 264.7 and C6 whole cell lysates; Human liver and heart tissue lysates; Mouse liver and heart tissue lysates; Rat liver and tissue lysates. IHC-P: Human liver and ovarian adenocarcinoma tissue; Mouse liver tissue; Rat liver tissue. Flow cyt: HepG2 and Hepa1-6 cells. IP: HepG2 whole cell lysate; Mouse liver tissue lysate.

  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab259967 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
IP 1/30.
WB 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 33 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/50.
  • Application notes
    Is unsuitable for ICC.

    • Target

    • Function

      NAD-dependent lysine demalonylase, desuccinylase and deglutarylase that specifically removes malonyl, succinyl and glutaryl groups on target proteins (PubMed:21908771, PubMed:22076378, PubMed:24703693). Activates CPS1 and contributes to the regulation of blood ammonia levels during prolonged fasting: acts by mediating desuccinylation and deglutarylation of CPS1, thereby increasing CPS1 activity in response to elevated NAD levels during fasting (PubMed:22076378, PubMed:24703693). Activates SOD1 by mediating its desuccinylation, leading to reduced reactive oxygen species (PubMed:24140062). Modulates ketogenesis through the desuccinylation and activation of HMGCS2 (By similarity). Has weak NAD-dependent protein deacetylase activity; however this activity may not be physiologically relevant in vivo. Can deacetylate cytochrome c (CYCS) and a number of other proteins in vitro such as UOX.

    • Tissue specificity

      Widely expressed.

    • Sequence similarities

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

    • Domain

      In contrast to class I sirtuins, class III sirtuins have only weak deacetylase activity. Difference in substrate specificity is probably due to a larger hydrophobic pocket with 2 residues (Tyr-102 and Arg-105) that bind to malonylated and succinylated substrates and define the specificity (PubMed:22076378).

    • Cellular localization

      Mitochondrion; Cytoplasm. Mitochondrion and Mitochondrion matrix. Mitochondrion intermembrane space. Cytoplasm, cytosol. Nucleus. Mainly mitochondrial. Also present extramitochondrially: a fraction is present in the cytosol and very small amounts are also detected in the nucleus.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • NAD dependent deacetylase sirtuin 5 antibody
      • NAD dependent lysine demalonylase and desuccinylase sirtuin 5 mitochondrial antibody
      • NAD dependent protein deacylase sirtuin 5 mitochondrial antibody
      • NAD-dependent protein deacylase sirtuin-5, mitochondrial antibody
      • Regulatory protein SIR2 homolog 5 antibody
      • Silent mating type information regulation 2 S.cerevisiae homolog 5 antibody
      • Sir2 like 5 antibody
      • SIR2-like protein 5 antibody
      • SIR2L5 antibody
      • SIR5_HUMAN antibody
      • Sirt5 antibody
      • Sirtuin 5 antibody
      • Sirtuin type 5 antibody
      see all

    Images

    • Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

      Lane 1 : Mouse liver tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : Rat liver tissue lysate
      Lane 4 : Rat heart tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

      Predicted band size: 33 kDa
      Observed band size: 30 kDa
      why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 48 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse liver (PMID 29491006). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      SIRT5 was immunoprecipitated from 0.35 mg Mouse liver tissue lysate with ab259967 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259967 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse liver tissue lysate 10 ug

      Lane 2: ab259967 IP in Mouse liver tissue lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259967 in mouse liver tissue lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes.

       

    • Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

      Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 2 : Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 4 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
      Lane 5 : Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate
      Lane 6 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 7 : C6 (rat glial tumor glial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

      Predicted band size: 33 kDa
      Observed band size: 30 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver (PMID 29491006). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      SIRT5 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab259967 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259967 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

      Lane 2: ab259967 IP in HepG2 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259967 in HepG2 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes.

       

    • Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Hepa1-6 (Mouse hepatoma epithelial cell) cells labelling SIRT5 with ab259967 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

      A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

      Lane 1 : Human liver tissue lysate
      Lane 2 : Human heart tissue lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

      Predicted band size: 33 kDa
      Observed band size: 30 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat liver (PMID 29491006). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling SIRT5 with ab259967 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

      A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

      Immunohistochemical analysis of paraffin-embedded Human ovarian adenocarcinoma tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human ovarian adenocarcinoma (PMID 31456942). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab259967? Please let us know so that we can cite the reference in this datasheet.

    ab259967 has not yet been referenced specifically in any publications.

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