Human NAD-dependent ADP-ribosyltransferase sirtuin-4 (SIRT4/SIR2L4) ELISA Kit

SIRT4 encodes a member of the sirtuin family of proteins which are homologs of the Sir2 gene in budding yeast. Members of the sirtuin family are characterized by a sirtuin core domain and grouped into four classes. The functions of human sirtuins have not yet been fully determined; however, yeast sirtuin proteins are known to regulate epigenetic gene silencing and suppress recombination of rDNA. Studies suggest that the human sirtuins may function as intracellular regulatory proteins with mono-ADP-ribosyltransferase activity. The protein encoded by this gene is included in class IV of the sirtuin family. SIRT4 is a mitochondrial ADP-ribosyltransferase that inhibits mitochondrial glutamate dehydrogenase 1 activity, thereby downregulating insulin secretion in response to amino acids.

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Human NAD-dependent ADP-ribosyltransferase sirtuin-4 (SIRT4/SIR2L4) ELISA Kit has high sensitivity and excellent specificity for detection of Human SIRT4. No significant cross-reactivity or interference between Human SIRT4 and analogues was observed.

This Human NAD-dependent ADP-ribosyltransferase sirtuin-4 (SIRT4/SIR2L4) ELISA Kit employs a two-site sandwich ELISA to quantitate SIRT4 in samples. An antibody specific for SIRT4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anySIRT4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for SIRT4 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of SIRT4 bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.