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Cell Biology Apoptosis Intracellular p53 Pathway
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RecombinantSimpleStep

Mouse SIRT1 ELISA Kit (ab206983)

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Other - Mouse SIRT1 ELISA Kit (ab206983)
  • Example of SIRT1 standard curve.
  • Interpolated concentrations of SIRT1 in mouse NIH3T3 extract and C2C12 extract based on a 1 mg/mL extract load.
  • Interpolated concentrations of SIRT1 in rat PC12 extract and human HEK293 extract based on a 1 mg/mL extract load.
  • Sandwich ELISA - Mouse SIRT1 ELISA Kit (ab206983)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 31 pg/ml
  • Range: 156.25 pg/ml - 10000 pg/ml
  • Sample type: Cell culture extracts, Tissue Extracts
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Mouse

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Overview

  • Product name

    Mouse SIRT1 ELISA Kit
    See all SIRT1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    NIH3T3 5 6.9%
    Inter-assay
    Sample n Mean SD CV%
    NIH3T3 3 4.8%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    31 pg/ml
  • Range

    156.25 pg/ml - 10000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Mouse brain extract 97 89% - 103%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Mouse SIRT1 ELISA Kit (ab206983) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of SIRT1 protein in cell culture extracts and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse SIRT1 with 31 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • 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. Sirtuin 1 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.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse SIRT1 Capture Antibody 1 x 600µl
    10X Mouse 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 5BR 1 x 6ml
    Mouse SIRT1 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • p53 Pathway
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • Homologous Recomb.
    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Other Nuclear Bodies
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • HDACs
    • Class III / Sir2 class
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Apoptosis marker and proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • DNA Damage ELISA kits
    • Metabolism
    • Types of disease
    • Obesity
  • Function

    NAD-dependent protein deacetylase that links transcriptional regulation directly to intracellular energetics and participates in the coordination of several separated cellular functions such as cell cycle, response to DNA damage, metobolism, apoptosis and autophagy. Can modulate chromatin function through deacetylation of histones and can promote alterations in the methylation of histones and DNA, leading to transcriptional repression. Deacetylates a broad range of transcription factors and coregulators, thereby regulating target gene expression positively and negatively. Serves as a sensor of the cytosolic ratio of NAD(+)/NADH which is altered by glucose deprivation and metabolic changes associated with caloric restriction. Is essential in skeletal muscle cell differentiation and in response to low nutrients mediates the inhibitory effect on skeletal myoblast differentiation which also involves 5'-AMP-activated protein kinase (AMPK) and nicotinamide phosphoribosyltransferase (NAMPT). 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 'Lys-266' of SUV39H1, leading to its activation. Inhibits skeletal muscle differentiation by deacetylating PCAF and MYOD1. Deacetylates H2A and 'Lys-26' of HIST1H1E. Deacetylates 'Lys-16' of histone H4 (in vitro). Involved in NR0B2/SHP corepression function through chromatin remodeling: Recruited to LRH1 target gene promoters by NR0B2/SHP thereby stimulating histone H3 and H4 deacetylation leading to transcriptional repression. Proposed to contribute to genomic integrity via positive regulation of telomere length; however, reports on localization to pericentromeric heterochromatin are conflicting. Proposed to play a role in constitutive heterochromatin (CH) formation and/or maintenance through regulation of the available pool of nuclear SUV39H1. Upon oxidative/metabolic stress decreases SUV39H1 degradation by inhibiting SUV39H1 polyubiquitination by MDM2. This increase in SUV39H1 levels enhances SUV39H1 turnover in CH, which in turn seems to accelerate renewal of the heterochromatin which correlates with greater genomic integrity during stress response. Deacetylates 'Lys-382' of p53/TP53 and impairs its ability to induce transcription-dependent proapoptotic program and modulate cell senescence. Deacetylates TAF1B and thereby represses rDNA transcription by the RNA polymerase I. Deacetylates MYC, promotes the association of MYC with MAX and decreases MYC stability leading to compromised transformational capability. Deacetylates FOXO3 in response to oxidative stress thereby increasing its ability to induce cell cycle arrest and resistance to oxidative stress but inhibiting FOXO3-mediated induction of apoptosis transcriptional activity; also leading to FOXO3 ubiquitination and protesomal degradation. Appears to have a similar effect on MLLT7/FOXO4 in regulation of transcriptional activity and apoptosis. Deacetylates DNMT1; thereby impairs DNMT1 methyltransferase-independent transcription repressor activity, modulates DNMT1 cell cycle regulatory function and DNMT1-mediated gene silencing. Deacetylates RELA/NF-kappa-B p65 thereby inhibiting its transactivating potential and augments apoptosis in response to TNF-alpha. Deacetylates HIF1A, KAT5/TIP60, RB1 and HIC1. Deacetylates FOXO1 resulting in its nuclear retention and enhancement of its transcriptional activity leading to increased gluconeogenesis in liver. Inhibits E2F1 transcriptional activity and apoptotic function, possibly by deacetylation. Involved in HES1- and HEY2-mediated transcriptional repression. In cooperation with MYCN seems to be involved in transcriptional repression of DUSP6/MAPK3 leading to MYCN stabilization by phosphorylation at 'Ser-62'. Deacetylates MEF2D. Required for antagonist-mediated transcription suppression of AR-dependent genes which may be linked to local deacetylation of histone H3. Represses HNF1A-mediated transcription. Required for the repression of ESRRG by CREBZF. Modulates AP-1 transcription factor activity. Deacetylates NR1H3 AND NR1H2 and deacetylation of NR1H3 at 'Lys-434' positively regulates transcription of NR1H3:RXR target genes, promotes NR1H3 proteosomal degradation and results in cholesterol efflux; a promoter clearing mechanism after reach round of transcription is proposed. Involved in lipid metabolism. Implicated in regulation of adipogenesis and fat mobilization in white adipocytes by repression of PPARG which probably involves association with NCOR1 and SMRT/NCOR2. Deacetylates ACSS2 leading to its activation, and HMGCS1. Involved in liver and muscle metabolism. Through deacteylation and activation of PPARGC1A is required to activate fatty acid oxidation in skeletel muscle under low-glucose conditions and is involved in glucose homeostasis. Involved in regulation of PPARA and fatty acid beta-oxidation in liver. Involved in positive regulation of insulin secretion in pancreatic beta cells in response to glucose; the function seems to imply transcriptional repression of UCP2. Proposed to deacetylate IRS2 thereby facilitating its insulin-induced tyrosine phosphorylation. Deacetylates SREBF1 isoform SREBP-1C thereby decreasing its stability and transactivation in lipogenic gene expression. Involved in DNA damage response by repressing genes which are involved in DNA repair, such as XPC and TP73, deacetylating XRCC6/Ku70, and faciliting recruitment of additional factors to sites of damaged DNA, such as SIRT1-deacetylated NBN can recruit ATM to initiate DNA repair and SIRT1-deacetylated XPA interacts with RPA2. Also involved in DNA repair of DNA double-strand breaks by homologous recombination and specifically single-strand annealing independently of XRCC6/Ku70 and NBN. Transcriptional suppression of XPC probably involves an E2F4:RBL2 suppressor complex and protein kinase B (AKT) signaling. Transcriptional suppression of TP73 probably involves E2F4 and PCAF. Deacetylates WRN thereby regulating its helicase and exonuclease activities and regulates WRN nuclear translocation in response to DNA damage. Deacetylates APEX1 at 'Lys-6' and 'Lys-7' and stimulates cellular AP endonuclease activity by promoting the association of APEX1 to XRCC1. Increases p53/TP53-mediated transcription-independent apoptosis by blocking nuclear translocation of cytoplasmic p53/TP53 and probably redirecting it to mitochondria. Deacetylates XRCC6/Ku70 at 'Lys-539' and 'Lys-542' causing it to sequester BAX away from mitochondria thereby inhibiting stress-induced apoptosis. Is involved in autophagy, presumably by deacetylating ATG5, ATG7 and MAP1LC3B/ATG8. Deacetylates AKT1 which leads to enhanced binding of AKT1 and PDK1 to PIP3 and promotes their activation. Proposed to play role in regulation of STK11/LBK1-dependent AMPK signaling pathways implicated in cellular senescence which seems to involve the regulation of the acetylation status of STK11/LBK1. Can deacetylate STK11/LBK1 and thereby increase its activity, cytoplasmic localization and association with STRAD; however, the relevance of such activity in normal cells is unclear. In endothelial cells is shown to inhibit STK11/LBK1 activity and to promote its degradation. Deacetylates SMAD7 at 'Lys-64' and 'Lys-70' thereby promoting its degradation. Deacetylates CIITA and augments its MHC class II transactivation and contributes to its stability. Deacteylates MECOM/EVI1. Deacetylates PML at 'Lys-487' and this deacetylation promotes PML control of PER2 nuclear localization. During the neurogenic transition, repress selective NOTCH1-target genes throug
    Isoform 2: Isoform 2 is shown to deacetylate 'Lys-382' of p53/TP53, however with lower activity than isoform 1. In combination, the two isoforms exert an additive effect. Isoform 2 regulates p53/TP53 expression and cellular stress response and is in turn repressed by p53/TP53 presenting a SIRT1 isoform-dependent auto-regulatory loop.
    (Microbial infection) In case of HIV-1 infection, interacts with and deacetylates the viral Tat protein. The viral Tat protein inhibits SIRT1 deacetylation activity toward RELA/NF-kappa-B p65, thereby potentiates its transcriptional activity and SIRT1 is proposed to contribute to T-cell hyperactivation during infection.
    SirtT1 75 kDa fragment: catalytically inactive 75SirT1 may be involved in regulation of apoptosis. May be involved in protecting chondrocytes from apoptotic death by associating with cytochrome C and interfering with apoptosome assembly.
  • Tissue specificity

    Widely expressed.
  • Sequence similarities

    Belongs to the sirtuin family. Class I subfamily.
    Contains 1 deacetylase sirtuin-type domain.
  • Post-translational
    modifications

    Methylated on multiple lysine residues; methylation is enhanced after DNA damage and is dispensable for deacetylase activity toward p53/TP53.
    Phosphorylated. Phosphorylated by STK4/MST1, resulting in inhibition of SIRT1-mediated p53/TP53 deacetylation. Phosphorylation by MAPK8/JNK1 at Ser-27, Ser-47, and Thr-530 leads to increased nuclear localization and enzymatic activity. Phosphorylation at Thr-530 by DYRK1A and DYRK3 activates deacetylase activity and promotes cell survival. Phosphorylation by mammalian target of rapamycin complex 1 (mTORC1) at Ser-47 inhibits deacetylation activity. Phosphorylated by CaMK2, leading to increased p53/TP53 and NF-kappa-B p65/RELA deacetylation activity (By similarity). Phosphorylation at Ser-27 implicating MAPK9 is linked to protein stability. There is some ambiguity for some phosphosites: Ser-159/Ser-162 and Thr-544/Ser-545.
    Proteolytically cleaved by cathepsin B upon TNF-alpha treatment to yield catalytic inactive but stable SirtT1 75 kDa fragment (75SirT1).
    S-nitrosylated by GAPDH, leading to inhibit the NAD-dependent protein deacetylase activity.
  • Cellular localization

    Cytoplasm. Mitochondrion and Nucleus, PML body. Cytoplasm. Nucleus. Recruited to the nuclear bodies via its interaction with PML (PubMed:12006491). Colocalized with APEX1 in the nucleus (PubMed:19934257). May be found in nucleolus, nuclear euchromatin, heterochromatin and inner membrane (PubMed:15469825). Shuttles between nucleus and cytoplasm (By similarity). Colocalizes in the nucleus with XBP1 isoform 2 (PubMed:20955178).
  • 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

    • 75SirT1
    • hSIR2
    • hSIRT1
    • HST2
    • HST2, S. cerevisiae, homolog of
    • NAD dependent deacetylase sirtuin 1
    • NAD dependent protein deacetylase sirtuin 1
    • NAD-dependent deacetylase sirtuin-1
    • OTTHUMP00000198111
    • OTTHUMP00000198112
    • Regulatory protein SIR2 homolog 1
    • SIR1_HUMAN
    • SIR2
    • SIR2 like 1
    • SIR2 like protein 1
    • SIR2, S.cerevisiae, homolog-like 1
    • SIR2-like protein 1
    • SIR2ALPHA
    • SIR2L1
    • Sirt1
    • SirtT1 75 kDa fragment
    • Sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae)
    • Sirtuin 1
    • Sirtuin type 1
    see all
  • Database links

    • Entrez Gene: 93759 Mouse
    • SwissProt: Q923E4 Mouse
    • Unigene: 351459 Mouse

    Associated products

    • Matched antibody pairs

      • Mouse SIRT1 Matched Antibody Pair Kit (ab213465)

    Images

    • Other - Mouse SIRT1 ELISA Kit (ab206983)
      Other - Mouse SIRT1 ELISA Kit (ab206983)

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Example of SIRT1 standard curve.
      Example of SIRT1 standard curve.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of SIRT1 in mouse NIH3T3 extract and C2C12 extract based on a 1 mg/mL extract load.
      Interpolated concentrations of SIRT1 in mouse NIH3T3 extract and C2C12 extract based on a 1 mg/mL extract load.

      The concentrations of SIRT1 were measured in duplicate and interpolated from the SIRT1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean SIRT1 concentration was determined to be 1,732 pg/mL in mouse NIH3T3 extract and 3,530 pg/mL in mouse C2C12 extract.

    • Interpolated concentrations of SIRT1 in rat PC12 extract and human HEK293 extract based on a 1 mg/mL extract load.
      Interpolated concentrations of SIRT1 in rat PC12 extract and human HEK293 extract based on a 1 mg/mL extract load.

      The concentrations of SIRT1 were measured in duplicate and interpolated from the SIRT1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean SIRT1 concentration was determined to be 14,454 pg/mL in rat PC12 extract and 2606 pg/mL in human HEK293 extract.

    • Sandwich ELISA - Mouse SIRT1 ELISA Kit (ab206983)
      Sandwich ELISA - Mouse SIRT1 ELISA Kit (ab206983)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (1)

    Publishing research using ab206983? Please let us know so that we can cite the reference in this datasheet.

    ab206983 has been referenced in 1 publication.

    • Velagapudi R  et al. AMPK and SIRT1 activation contribute to inhibition of neuroinflammation by thymoquinone in BV2 microglia. Mol Cell Biochem 435:149-162 (2017). PubMed: 28551846

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Question

    I am interested in using this Sirt1 elisa to quantify the amount of Sirt1 in rat brain lysates. Can you confirm that this "mouse Sirt1 elisa" is compatible with rat tissue. Also, do you have any information as to how much protein to load for homogenates or any references where someone has used this elisa for brain homogenates.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 18 2015

    Answer

    Thank you for your interest in the SIRT1 SimpleStep® ELISA, ab206983. Rat reactivity was assessed with a test against PC12 lysates and the data is in figure 2 of the protocol booklet. The booklet is linked on the online datasheet near the upper right of the online datasheet.

    This is a fairly new assay which we added to the catalogue about a month ago, so we do not have data for brain homogenates from in-house tests or customers or references yet. (If you do decide to try the kit, I encourage you to send a review of your results, positive or negative, via our https://www.abcam.com/content/what-is-an-abreview program).

    Page 18 of the protocol has some data for various dilutions of mouse brain homogenate (MBH) starting with 250ug/mL total protein. The data shows that a range of protein concentrations from 250ug/mL to an 8-fold dilution, 31ug/mL, all gave OD450 values within the range of the standards.

    Read More

    Tom Ruyle

    Abcam Scientific Support

    Answered on Sep 18 2015

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