RNase A, DNase and protease-free
Thermo Scientific RNase A, DNase and protease-free is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues (see Figure 1 in Supporting Data).
It cleaves the phosphodiester bond between the 5'-ribose of a nucleotide and the phosphate group attached to the 3'-ribose of an adjacent pyrimidine nucleotide. The resulting 2', 3'-cyclic phosphate is hydrolyzed to the corresponding 3'-nucleoside phosphate (see References 1, 2).
- RNase A is free of DNase activity. It is not necessary to heat it before use.
- Plasmid and genomic DNA preparation (see References 3, 4)
- Removal of RNA from recombinant protein preparations
- Ribonuclease protection assays. Used in conjunction with RNase T1 (see Reference 3)
- Mapping single-base mutations in DNA or RNA (see References 5, 6)
Recommended concentration of RNase A is 1 to 100 µg/mL depending on the application. The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 to 100 mM NaCl), RNase A cleaves single-stranded and double-stranded RNA as well the RNA strand in RNA-DNA hybrids. However, at NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves single-stranded RNA (see Reference 9).
Protein concentration is determined by measuring the absorbance at 278 nm using the molar absorption coefficient at 9800 M-1 cm-1 (7).
|Definition of Activity Unit||
|Inactivation||Not inactivated by heating. Reliably removed by spin column or phenol/chloroform extraction.|
|Inhibition||Inhibitors: the most potent inhibitor is a mamalian ribonuclease inhibitor, e.g., <_a20_href3d_22_2f_general-reagents-and-accessories2f_ribolock-rnase-inhibitor2f_3e_ribolock20_rnase20_inhibitor3c_2f_a3e_.20_other20_inhibitors3a_20_uridine20_227_2c_20_327_-cyclic20_vanadate2c_20_527_-diphosphoadenosine20_327_-phosphate20_and20_527_-diphosphoadenosine20_227_-phosphate20_28_229_2c_20_sds2c_20_diethyl20_pyrocarbonate2c_20_4c2a0_m20_guanidinium20_thiocyanate20_plus20_0.1c2a0_m20_227_-mercaptoethanol2c_20_and20_heavy20_metal20_ions.>|
|Molecular Weight||13.7 kDa monomer|
|Specific Activity||>5000 U/mg protein (>100 Kunitz units/mg protein)|
|Storage Buffer||The enzyme is supplied in 50 mM Tris-HCl (pH 7.4) and 50% (v/v) glycerol.|
|Storage Condition||-20 C|
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