Description :
Deoxyribonuclease I (DNAse I) is an endonuclease that cleaves DNA phosphodiester bonds yielding 5′-phosphorylated and 3′-hydroxylated oligonucleotides. DNAse I targets single-stranded DNA, double-stranded DNA, and chromatin in a non-specific manner.As an important player in cellular waste management, DNAse I is normally secreted extracellularly to clear the system from circulating cell-free DNA, foreign DNA from food digestion or potential pathogens, and endogenous chromosomal DNA from apoptotic and necrotic cells. Abnormal DNAse I activity occurs in association with a variety of cancers and auto-immune illnesses that exhibit elevated levels of cell-free DNA. Furthermore, DNAse I has been therapeutically used in cystic fibrosis patients to degrade DNA and reduce sputum viscosity. DNAse I Activity Fluorometric Assay Kit allows for quantitative evaluation of DNAse I activity of purified enzymes and their inhibitors as well as comparative examination of DNAse I activity in biological samples. Enzyme activity is detected upon cleavage of a DNA Probe, which yields a fluorescent DNA product measured at Ex/Em = 651/681 nm. The limit of quantification (L.O.Q) is 178 fmoles of DNA probe cleaved per minute per ml.
Applications :
Measurement of DNAse I activity ofpurified proteins;
Quantitative analysis of DNAse I mutants and inhibitors;
Comparative examination of DNAse I activity in biological samples
Storage :
Store kit at -20°C, protected from light. Briefly centrifuge small vials prior to opening. Upon re-suspension, aliquot DNA Probe and DNAse I Positive control to avoid repeated freeze-thaw cycles. Read entire protocol before performing the assay.
• 10X DNAse I Assay Buffer: Store at 4 ºC. Warm to 37 ºC temperature before use.
• DNA Probe: Reconstitute with 220 μl of DNA Probe Re-suspension Buffer. Aliquot and store at -20ºC. Avoid multiple freeze-thaw cycles.
• DNA Probe Re-suspension Buffer: Ready to use. Store at RT.
• DNAse I Positive Control: Reconstitute with 220 μl of Positive Control Re-suspension Buffer. Aliquot and store at -20°C.
• Positive Control Re-suspension Buffer: Ready to use. Store at -20°C.
• Molecular Biology Grade Water: Ready to use. Store at RT.
Kit Components :
10X DNAse I Assay Buffer, 1.1 ml
DNA Probe, 1 vial
DNA Probe Re-suspension Buffer, 250 μl
DNAse I Positive Control, 1 vial
Positive Control Re-suspension Buffer, 1 ml
Molecular Biology Grade Water, 25 ml
Compatible Sample Types :
Purified Protein, serum.