Recombinant Micrococcal Nuclease

 

Catalog # : EPX-016-REP

 

Source : Staphylococcus aureus

 

Expressed in : E. Coli

 

Quantity : 500 UN at 0.5U /ul

 

 

 

Background:

 

Micrococcal nuclease or MNase is a 16.9 kDa endonuclease derived from Staphylococcus aureus (Gene ID: 3238436).  It is purified from an E. coli strain expressing an N-terminal 6XHIS tagged micrococcal nuclease.. Purified protein exhibit an strong endonuclease activity against single-stranded, double-stranded, circular and linear nucleic acids. The enzyme is active in the pH range of 7.0 - 10.0, with optimal activity at pH 9.2 for both RNA and DNA substrates. The rate of cleavage is 30 times greater at the 5' side of A or T than at G or C and results in the production of mononucleotides and oligonucleotides with terminal 3’-phosphates

 

 

Protein details:

 

Micrococcal Nuclease (MNase) is suitable for removing nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments. The enzyme activity is strictly dependent on Ca2+ and the pH optimum varies according to Ca2+ concentration. The enzyme is therefore easily inactivated by EGTA or EDTA. Purified Micrococcal Nuclease is formulated in a storage buffer containing 20mM Tris-Cl pH 8.0, 50mM NaCl, 1mM DTT and 50% glycerol.



Unit Definition:

 

One unit will produce 1.0 μmole of acid soluble polynucleotides from native DNA per min at pH 8 at 37 C, based on EM/260 = 10,000 for the mixed nucleotides.

 

 

Quality control:

 

Purified recombinant micrococcal nuclease is evaluated by 12% SDS PAGE mini-gel for protein purity and by spectrometry and 1% agarose gel for nuclease activity (Figure 1, right and left panels).

 

 

Application Notes:

 

For research use only.

 

 

Storage:

 

-20C

 

 

Guarantee:

 

Products guaranteed stable for 2 years from date of receipt when stored properly.

 

 

Purity:

 

>98% purity by SDS PAGE.

 

 

Protein sequences:

H2A (Theoretical Mw: 14091.48)

MSGRGKQGGKARAKAKTRSSRAGLQFPVGRVHRLLRKGNYAERVGAGA

PVYLAAVLEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRNDEELNK

LLGKVTIAQGGVLPNIQAVLLPKKTESHHKAKGK

 

H2B (Theoretical Mw: 13904.17)

MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKVLKQ

VHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQ

TAVRLLLPGELAKHAVSEGTKAVTKYTSAK

 

H3.1 (Theoretical Mw: 15404.09)

MARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVKKPHRYRPGTVALR

EIRRYQKSTELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEACEAY

LVGLFEDTNLCAIHAKRVTIMPKDIQLARRIRGERA

 

H4 (Theoretical Mw: 11367.34)

MSGRGKGGKGLGKGGAKRHRKVLRDNIQGITKPAIRRLARRGGVKRIS

GLIYEETRGVLKVFLENVIRDAVTYTEHAKRKTVTAMDVVYALKRQGR

TLYGFGG

 

References:

1. Van Holde, K. E. (1989) Chromatin, 1-497.

2. Luger et al., (1997) Nature, 389(6648):251-60..

 

 

 


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