Overview

  • Product nameSIRT1 Human ELISA KitSee all SIRT1 kits ...
  • Detection methodColorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 8 7%
  • Tests
    1 x 96 well plate
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeSandwich (quantitative)
  • Sensitivity
    8 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 106 98% - 112%

  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse, Rat
  • Product overview

    ab123457 SIRT1 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human SIRT1 in cell and tissue lysates. The assay employs an antibody specific for Human SIRT1 protein coated onto well plate strips.

    Standards and samples are pipetted into the wells and analyte present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti- SIRT1 primary detector antibody is added. After washing away unbound primary detector antibody, HRP-conjugated secondary detector antibody specific for the primary detector antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of analyte bound. The developing blue color is measured at 600 nm.

    Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.

     

    Species– human. Rat, mouse are not suitable with this kit, other species are untested.

  • Notes

    Typical working ranges:

    Tissue extract (e.g. Heart): None detected

    Whole cultured cell extract: 0.060 - 1 mg/mL

    Human SIRT1 standard: 0.008 - 0.5 µg/mL

    Store all components at 4°C. This kit is stable for at least 6 months from receipt. After reconstitution the standard should be stored at -80°C. Unused microplate strips should be returned to the pouch containing the desiccant and resealed.

  • Tested applicationsSandwich ELISA more details
  • PlatformMicroplate

Properties

  • FunctionNAD-dependent protein deacetylase, which regulates processes such as apoptosis and muscle differentiation by deacetylating key proteins. Deacetylates 'Lys-382' of p53/TP53 and impairs its ability to induce proapoptotic program and modulate cell senescence. Deacetylates TAF1B and thereby represses rDNA transcription by the RNA polymerase I. Deacetylates 'Lys-266' of SUV39H1, leading to its activation. Deacetylates 'Lys-26' of HIST1H1E. Involved in HES1- and HEY2-mediated transcriptional repression. Inhibits skeletal muscle differentiation by deacetylating PCAF and MYOD1. May serve as a sensor of the cytosolic ratio of NAD(+)/NADH, which is essential in skeletal muscle cell differentiation. Deacetylates 'Lys-16' of histone H4 (in vitro). Component of the eNoSC (energy-dependent nucleolar silencing) complex, a complex that mediates silencing of rDNA in response to intracellular energy status and acts by recruiting histone-modifying enzymes. The eNoSC complex is able to sense the energy status of cell: upon glucose starvation, elevation of NAD(+)/NADP(+) ratio activates SIRT1, leading to histone H3 deacetylation followed by dimethylation of H3 at 'Lys-9' (H3K9me2) by SUV39H1 and the formation of silent chromatin in the rDNA locus. Deacetylates H2A. In case of HIV-1 infection, interacts with and deacetylates the viral Tat protein. Deacetylates APEX1 at 'Lys-6' and 'Lys-7'. Stimulates cellular AP endonuclease activity by promoting the association of APEX1 to XRCC1.
  • Tissue specificityWidely expressed.
  • Sequence similaritiesBelongs to the sirtuin family.
    Contains 1 deacetylase sirtuin-type domain.
  • Post-translational
    modifications
    S-nitrosylated by GAPDH, leading to inhibit the NAD-dependent protein deacetylase activity.
  • Cellular localizationNucleus > PML body. Recruited to the nuclear bodies via its interaction with PML. Colocalized with APEX1 in the nucleus. May be found in nucleolus, nuclear euchromatin, heterochromatin and inner membrane.
  • Target information above from: UniProt accession Q96EB6 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names
    • hSIR2
    • hSIRT1
    • NAD-dependent deacetylase sirtuin-1
    • Regulatory protein SIR2 homolog 1
    • SIR2-like protein 1
    • SIR2L1
    • Sirt1
    • SIRT1_HUMAN
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab123457 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Sandwich ELISA Use at an assay dependent concentration.

SIRT1 Human ELISA Kit images

  • Example standard curve. Serially diluted recombinant SIRT1 in the working range of the assay.
  • Example of serially titrated HeLa extract in the assay working range.
  • The expression level of SIRT1 varies by cell type. Four cell types are analyzed by SIRT1 expression level relative to the Human HepG2 cell line. This relative relationship is confirmed by Western blotting using antibody ab13533.
  • The SIRT1 detector antibody (ab110304) in this ELISA is available as standalone antibody. In this image Immunocytochemistry of the antibody (ab110304) from this ELISA used to stain Human HDFn cells.

Protocols

References for SIRT1 Human ELISA Kit (ab123457)

ab123457 has not yet been referenced specifically in any publications.

Product Wall

Yes, the SIRT1 ELISA ab123457 has been tested in 10% serum and yielded an average spike recovery of 106% (page 18 of booklet). The minimum detectable dose (MDD) for the kit is 8ng/mL.

I recommend the SIRT1 Human SimpleStep™ ELISA Kit (ab171573...

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The optimal number of cells is 2x107/mL.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for confirming that and sorry for the delay in getting back to you.

I am sorry butwe do not currentlyhave aSIRT1 ELISA kit specificallydesigned to recognize the mouse protein.Thehuman SIRT1 protein, for which the kit ab123457 is des...

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Thank you fro contacting us.


The reference to “recovery” in the protocol book of SIRT1 Human ELISA Kit (ab123457) might also be called “interference”. The point of this experiment is to validate the ELISA kit to ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"