Mouse SIRT1 ELISA Kit (ab206983)
- Datasheet
- References (1)
- Protocols
Overview
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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
Abcam’s SIRT1 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of SIRT1 protein in mouse cell and tissue extracts.
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.
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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.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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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
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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
modificationsS-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
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Alternative names
- hSIR2
- hSIRT1
- NAD-dependent deacetylase sirtuin-1
see all -
Database links
- Entrez Gene: 93759 Mouse
- SwissProt: Q923E4 Mouse
Associated products
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Matched antibody pairs
Images
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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.
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Example of SIRT1 standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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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.
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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.
Protocols
References
This product has been referenced in:
- 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). Read more (PubMed: 28551846) »
