NucA nuclease Antibody
NucA nuclease Antibody Summary
Met1-Asn266
Accession # P13717
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of NucA nuclease by Western Blot. Western blot shows recombinant NucA nuclease. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-NucA nuclease Monoclonal Antibody (Catalog # MAB10912) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for NucA nuclease at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
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Detection of NucA nuclease by Simple WesternTM. Simple Western lane view shows recombinant NucA nuclease protein, loaded at 0.2 mg/mL. A specific band was detected for NucA nuclease at approximately 35 kDa (as indicated) using 10 µg/mL of Rabbit Anti-NucA nuclease Monoclonal Antibody (Catalog # MAB10912). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: NucA nuclease
Serratia marcescens endonuclease, also known as Benzonase, is a non-specific nuclease that degrades both single- and double-stranded nucleic acids, including DNA and RNA, but exhibits no proteolytic activity (1); therefore, it is ideal for the removal of nucleic acid contaminants from protein samples and applications where complete digestion of nucleic acids is desirable. NucA (Benzonase) is also commonly used in bioprocessing applications to reduce viscosity of samples caused by genomic DNA. The optimum pH for enzyme activity is found to be 8.0-9.2. It hydrolyzes internal phosphodiester bonds between nucleotides in nucleic acids to produce 5'-monophosphate oligonucleotides of 3-8 bases in length (2). The active enzyme is a homodimer with two disulfide bonds in each monomer that are crucial to the enzyme activity and stability (3). The absolute activity of the recombinant enzyme is measured by a phosphatase coupled assay (4), where the 5'-phosphate of oligosaccharides generated by the enzyme is further released by non-specific alkaline phosphatase and quantitated by Malachite reagents (5).
- Benedik, MJ and Strych, U. (1998) FEMS Microbiol Lett. 165:1.
- Nestle, M, et al. (1999) J. Biol. Chem. 274:825.
- Ball, T.K. et al. (1992) Nucleic Acids Res. 20:4971.
- Wu, Z.L. et al. (2011) Glycobiology 21:727.
- Van Veldhoven, P.P. and G.P. Mannaerts (1987) Anal. Biochem. 161:45.
Product Datasheets
Product Specific Notices
Benzonase is a registered trademark of Merck KGaA Corp.FAQs
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