제품 설명
Endonuclease IV는 endonuclease IV 유전자를 E. coli에서 발현시킨 재조합 단백질입니다. Endonuclease로 명명되었지만 endonuclease와 exonuclease 활성을 모두 가지고 있습니다. 이 효소는 DNA에서 염기만이 제거된 apurinic 혹은 apyrimidinic (AP)부위에 작용하여 AP 부위의 5'-쪽 phosphodiester 결합을 절단하여 3'-OH와 5'-phosphate 를 가진 nick을 만듭니다. 이러한 활성은 DNA base excision repair에 필수적인 역할을 합니다. 또한 3'→5' exonucleas의 활성은 3' 말단이 오목하게 들어간 (recessed) 형태의 DNA 기질을 선호합니다.
제품 특징
- Isolated from a recombinant source
- Reaction temperature: 37℃
- Heat inactivation: 85℃ for 20 min
- Standard reaction: 1X Endonuclease IV Buffer, incubate at 37°C.
제품 응용
- Studies of DNA damage and repair
- Single cell electrophoresis (comet assay)
- DNA structure research
- Alkaline unwinding
제품 구성품
- Endonuclease IV (E. coli)
- 10X Endonuclease IV Buffer
- Sterile water
Unit 정의
One unit is defined as the amount of enzyme required to cleave 1 pmol of a 34-mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 µl in 1 hour at 37°C.
(* An AP site is created by treating 10 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 min at 37℃.)
Storage Buffer
10 mM Tris-HCl, 250 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.15% Triton X-100, 200 ug/ml BSA, 50% Glycerol, pH 7.4
10X Reaction Buffer
500 mM Tris-HCl, 1000 mM NaCl, 100 mM MgCl2, 10 mM DTT, pH 7.9
Figure 1. Cleavage of DNA containing an AP site by Endonuclease IV
20 pmol of 5’ end-labeled dsDNA substrates were incubated with Endonuclease IV at 37˚C. Cleaved DNA were analyzed on 15% Urea polyacrylamide gels in 0.5X TBE.
