Ordering Information
- : 17-10090
- : 96 assays
Product Images
Overview of SIRTainty Assay:
A hallmark of sirtuin enzymes is the generation of nicotinamide due to the cleavage of NAD+ during sirtuin m...
Sensitive Detection of Multiple Sirtuin Enzymes:
Sirtuins 1, 2, and 3 at a range of concentrations were incubated with 25 µM acetylated pe...
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SIRTainty™ Class III HDAC Assay
Description:
SIRTainty™ Class III HDAC Assay
Trade Name:
Upstate (Millipore)
Qty/Pk:
96 assays
Product Overview:
Features and Benefits
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.
- 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.
Key Applications:
Enzyme Assay
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.
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
Presentation:
Single 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.
Storage Conditions:
Maintain 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.
Promotional Text:
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)
Product Name:
SIRTainty™ Class III HDAC Assay
Materials Required but Not Delivered:
96-well microtiter plate reader with fluorescence readout
Plate shaker
Plate shaker