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17-10090 | SIRTainty™ Class III HDAC Assay

17-10090
96 assays  
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      Overview

      Replacement Information

      Special Offers

      Calbiochem® Brand Inhibitors and Enzymes
      • SIRT1, GST-Fusion, Human, Recombinant, E. coli (cat# 524743)
      • SIRT2, His•Tag®, Human, Recombinant, E. coli (cat# 524744)
      • SIRT3, GST Fusion, Human, Recombinant, E. coli (cat# 524745)
      • Suramin, Sodium Salt (cat# 574625)
      • SIRT1/2 Inhibitor IV, Cambinol (cat# 566323)
      • SIRT1/2 Inhibitor VII (cat# 566327)
      • SIRT1/2 Inhibitor VIII, Salermide (cat# 566330)
      • SIRT1 Inhibitor III (cat# 566322)
      • SIRT1 Inhibitor IV, (S)-35 (cat# 566325)
      • SIRT2 Inhibitor, AGK2 (cat# 566324)
      • SIRT2 Inhibitor II, AK-1 (cat# 566331)
      • Sirtinol (cat# 566320)
      • InSolution™ Sirtinol (cat# 566321)


      Key Specifications Table

      Key Applications
      EA
      Description
      Catalogue Number17-10090
      Trade Name
      • SIRTainty
      • Upstate
      DescriptionSIRTainty™ Class III HDAC Assay
      OverviewFeatures and Benefits
      • Sensitive Detection – assay design enables detection of all known sirtuin family members. Extensively validiated for sirtuins 1-3.
      • Unparalleled Flexibility – use of unlabeled substrates enables assay customization and the analysis of a variety of peptide substrates.
      • High Throughput – 96-well plate format ideal for inhibitor/activator assays and the study of enzyme mechanisms. Assay design can be adapted to 384-well plates.
      • Easy-to-use – simple protocol, homogeneous, no wash format.
      • Reliable Chemistry – unlabeled peptide substrate avoids detection artifacts due compounds such as resveratrol.
      • Rapid Results –easy two step assay with minimal hands on time allows you to go from sample to results in about 90 minutes.


      The SIRTainty™ Class III HDAC Assay utilizes a novel patent pending technology for the sensitive detection of all known sirtuin family members. This easy to perform assay employs nicotinamidase to measure nicotinamide generated upon cleavage of NAD+ during sirtuin mediated deacetylation of a substrate providing a direct assessment of the activity of class III HDAC enzymes.

      Unlike conventional assays that are dependent upon a single pre-labeled fluorescently tagged substrate, the SIRTainty Class III HDAC Assay employs untagged acetylated peptide substrate for the detection sirtuin activity. This approach not only enables unparalleled flexibility in the choice of sirtuin isoform and peptide substrate, but also eliminates the potential for artifacts due to the use of artificial substrates containing bulky fluorophores.
      Alternate Names
      • Sirtuin assay
      • Sirtuin HDAC assay
      • SIRT assay
      • SIRT HDAC assay
      Background InformationSirtuin, or Sir2 proteins, are a class of proteins that possess either mono-ADP-ribosyltransferase or deacylase activity, including deacetylase, desuccinylase, demalonylase, demyristoylase and depalmitoylase activity. Sirtuins have been implicated in influencing a wide range of cellular processes including aging, transcription, apoptosis, inflammation, stress resistance, as well as energy efficiency and alertness during low-calorie situations. Sirtuins can also control circadian clocks and mitochondrial biogenesis. The sirtuin-mediated deacetylation reaction couples lysine deacetylation to NAD hydrolysis. This hydrolysis yields O-acetyl-ADP-ribose, the deacetylated substrate and nicotinamide, itself an inhibitor of sirtuin activity. The dependence of sirtuins on NAD links their enzymatic activity directly to the energy status of the cell via the cellular NAD:NADH ratio, the absolute levels of NAD, NADH or nicotinamide or a combination of these variables.
      Materials Required but Not Delivered96-well microtiter plate reader with fluorescence readout
      Plate shaker
      References
      Product Information
      Components
      • Black 96 well plate and covers
      • SIRTaintyTM Assay Buffer
      • Acetylated Peptide Substrate
      • β-NAD
      • Nicotinamide (NAM)
      • Recombinant Sirt1 Enzyme
      • Recombinant Nicotinamidase Enzyme
      • Suramin (Sirt Inhibitor)
      • Developer Reagent
      PresentationSingle box containing all key reagents for evaluting the activity of class III histone deacetylases (sirtuins). Assay format is ideal for screening activators and inhibitors of all known sirtuin enzymes (sirts 1-7). Kit includes, sirtuin inhibitor, nicotinamide standard for quantitative analysis, and recombinant Sirt1 enzyme sufficient for proccessing 96 samples using 1.5 units of enzyme per assay.
      Applications
      ApplicationClass III HDAC assay used for the detection sirtuin activity levels. Assay employs nicotinamidase to measure nicotinamide generated upon cleavage of NAD+ during sirtuin mediated deacetylation of a substrate providing a direct assessment of the activity of class III HDAC enzymes.
      Key Applications
      • Enzyme Assay
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain the unopened kit at -80ºC. Plates and covers can be stored at room temperature (18-25º C). Unused components should be stored at -80ºC. When stored properly this kit is good for up to 6 months after date of receipt. Avoid repeated freeze/thaw cycles, aliquot if necessary.
      Packaging Information
      Material Size96 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      References

      Reference overviewPub Med ID
      Evidence for a common mechanism of SIRT1 regulation by allosteric activators.
      Hubbard, Basil P, et al.
      Science, 339: 1216-9 (2013) 2013

      Show Abstract
      23471411 23471411

      Brochure

      Title
      Hallmarks of Aging (EMD)
      New Products: Volume 3, 2012
      Product Selection Guide - Antibodies, small molecule inhibitors, kits, assays and proteins for signaling research.

      Technical Info

      Title
      96 well Template 2 Up
      96 well Template Single
      Epigenetics Application Note: A Novel, Substrate-Agnostic Class III HDAC Activity Assay

      Data Sheet

      Title
      SIRTainty™ Class III HDAC Assay

      User Guides

      Title
      SIRTAINTY™ CLASS III HDAC ASSAY

      Newsletters / Publications

      Title
      Cellutions - The Newsletter for Cell Biology Researchers Vol 3:2012