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| Product Name | Endonuclease IV (10 U/μL) |
| Catalog No. | IVDRM-0237 |
| Description | Endonuclease IV, also known as Nfo, primarily recognizes apurinic/apyrimidinic (AP) sites in DNA. It cleaves the phosphodiester bond on the 5' side of the AP site, generating a 3'-hydroxyl terminus and a 5'-deoxyribose phosphate (dRP) terminus. In addition, the enzyme possesses 3'-diesterase activity, releasing phosphoglycolaldehyde, intact deoxyribose-5-phosphate, and inorganic phosphate from the 3' terminus of DNA. Typical applications include hydrolysis of AP sites to create nicks that provide critical initiation sites for DNA repair in FFPE sample restoration, single-cell gel electrophoresis (comet assay), alkaline elution, and alkaline unwinding assays. |
| Intended Use | Raw material enzyme for DNA damage and repair applications, including FFPE sample DNA repair (generating nicks at AP sites as initiation sites for repair), single-cell gel electrophoresis (comet assay), alkaline elution, and alkaline unwinding assays. |
| Principle / Technology | Endonuclease IV (Nfo) specifically recognizes apurinic/apyrimidinic (AP) sites in DNA and cleaves the phosphodiester bond 5' to the lesion, producing a 3'-OH and a 5'-dRP terminus. Its intrinsic 3'-diesterase activity removes 3' blocking groups such as phosphoglycolaldehyde, deoxyribose-5-phosphate, and phosphate from DNA 3' ends. |
| Performance Range / Specifications | - Free of non-specific DNase contamination. - Host (E. coli) genomic DNA residue < 0.05 copies/10 U. |
| Reaction Conditions / Protocol | Typical reaction at 37 °C. Quality control assay example: 100 U of enzyme incubated with 0.5 μg λDNA-HindIII digest in a 20 uL reaction at 37 °C for 16 h shows no change in DNA banding pattern by agarose gel electrophoresis, confirming the absence of non-specific DNase activity. |
| Storage Conditions | Store at -25 °C to -15 °C. |
| Shelf Life | 1 year |
| Package Specifications | 1,000 U 5,000 U |
| Quality Control | - Non-specific DNase: not detected (no change in λDNA-HindIII banding pattern after incubation of 100 U enzyme with 0.5 μg substrate at 37 °C for 16 h). - Host (E. coli) genomic DNA residue: < 0.05 copies/10 U. |
| Purity | Free of detectable non-specific DNase contamination. |
| Concentration | 10 U/uL |
| Source / Origin | Recombinant enzyme expressed in E. coli. |
| Expiration Date / Stability | Stable for 1 year when stored at -25 °C to -15 °C. |
| Application Notes / Precautions | - In FFPE sample repair workflows, the enzyme hydrolyzes AP sites to generate nicks, providing critical initiation sites for subsequent DNA repair. - Suitable for single-cell gel electrophoresis (comet assay), alkaline elution, and alkaline unwinding applications. - The generated 3'-hydroxyl termini are extendable by DNA polymerases, facilitating downstream repair or labeling reactions. |
| Recognition Site | Apurinic/apyrimidinic (AP) sites in DNA; cleaves the phosphodiester bond immediately 5' to the AP site. |
For research use only, not for clinical use.
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