Overview

  • Product name
    Human SIRT1 ELISA Kit
    See all SIRT1 kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 5 2%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 4.1%
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    132 pg/ml
  • Range
    0.63 ng/ml - 40 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture media 94 90% - 101%
    Fetal Bovine Serum 84 83% - 85%
    Bovine Serum Albumin 107 106% - 107%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    Abcam’s SIRT1/sirtuin1 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of SIRT1 protein in Human cell samples.

    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm.Optionally,instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

     

  • Notes

    SIRT1 - silent mating type information regulation 2 homolog (homolog of yeast Sir2) – encodes a member of the sirtuins family of deacetylases. The sirtuin 1 protein (gene SIRT1) is responsible for epigenetic gene silencing after recruitment to the nucleus. Sirtuin1 deactylates proteins, including histones, in a wide (and growing) variety of processes in apoptosis and senescence, muscle differentiation and may serve as a cytosolic NAD+/NADH sensor. This enzyme may also regulate the circadian clock of the cell in response to metabolic conditions. SIRT1 is inhibited by nicotinamide and may be activated by resveratrol, a component of red wine. Resveratrol may participate in activation of sirtuin proteins, and may therefore participate in an extended lifespan as it has been observed in yeast.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human SIRT1 Capture Antibody 1 x 600µl
    10X Human SIRT1 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5B 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Human SIRT1 Lyophilized Recombinant Protein 2 x 50ng
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas
  • Function
    NAD-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 specificity
    Widely expressed.
  • Sequence similarities
    Belongs 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 localization
    Nucleus > 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.
  • 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 ab171573 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Example SIRT1 standard curve. Background-subtracted data values (mean +/- SD) are graphed.

  • Titration of HeLa extract within the working range of the assay. Background subtracted data from duplicate measurements are plotted.

  • Comparison of SIRT1 expression in different cell lines. Background subtracted data from duplicate measurements are plotted. Note the relative lower expression of SIRT1 in the A431 cell line.

  • Interpolated values of sirtuin1 are plotted for the indicated cell lines based on a lysate load of 125 µg/mL. Western blot using the detector sirtuin1 antibody on the same lysates quantified above.

  • Immunocytochemical staining of Human HDFn cells using the SIRT1 detector antibody in this kit (ab110304). The target protein locates mainly in the nucleus.

Protocols

References

This product has been referenced in:
  • Bartoli-Leonard F  et al. Suppression of SIRT1 in Diabetic Conditions Induces Osteogenic Differentiation of Human Vascular Smooth Muscle Cells via RUNX2 Signalling. Sci Rep 9:878 (2019). Read more (PubMed: 30696833) »
  • Saini JS  et al. Nicotinamide Ameliorates Disease Phenotypes in a Human iPSC Model of Age-Related Macular Degeneration. Cell Stem Cell N/A:N/A (2017). Sandwich ELISA ; Human . Read more (PubMed: 28132833) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Suitable for human serum

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We used the sirt-1 elisa kit for quantification of sirt-1 levels in human serum. Samples were used as 1:2 dilutions in the PTR buffer provided (1 x conc.). After correcting for the dilution factor, sirt-1 concentrations obtained ranged between 1.5 and 15 ng/mL.

Christine Moelzer

Verified customer

Submitted Dec 08 2015

SIRT1 protein levels in differente cell lines

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We used the kit to quantify SIRT1 levels in different cell lines (fig. B) and used the purified SIRT1 protein for normalization (fig. A) via standard curve. The assay was performed as described in the manual. BCA assay was performed to quantify protein concentrations.

Mr. Christian Marx

Verified customer

Submitted Jan 13 2014

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