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RNase A

Code
A2760
CAS
9001-99-4
Molar mass
~13700 g/mol

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Box prices only valid with purchase of full box.

code packaging size price per unit box price per unit
Code & packaging Price per piece
A2760,0100
code
A2760,0100
packaging size
100 mg
price per unit
single 32,00€
box price per unit
27,20€x 6 units
A2760,0500
code
A2760,0500
packaging size
500 mg
price per unit
single 100,70€
box price per unit
A2760,1000
code
A2760,1000
packaging size
1 g
price per unit
single 169,30€
box price per unit
Physical Description:
Solid
Product Code:
A2760
Product Name:
RNase A
Short Description:
delivery form: salt-free, freeze dried
Specifications:
Assay: approx. 70 %
Activity: min. 70 U/mg (Kunitz)
Comment:
Ribonuclease A (RNase A) is an endoribonuclease, that specifically cleaves single-stranded RNA 3' to pyrimidine residues (cytosine, uracil). Thereby, it generates pyrimidine-3'-phosphate or oligonucleotides with terminal pyrimidine-3'-phosphates. The pH-optimum is in the range of 7.0 - 7.5. RNase A is used for the purification of RNA-free DNA, for the removal of non-hybridized regions of RNA \: DNA-hybrides or as a molecular weight marker. The enzyme is inhibited by diethyl pyrocarbonate (DEPC), guanidinium salts (4 M GuaSCN), β-mercaptoethanol, heavy metals, vanadyl-ribonucleoside-complexes, RNase-inhibitor from human placenta and competitively by DNA, respectively. Regarding the latter, the effect of denatured DNA is higher than by native nucleic acids. Nevertheless, RNase A is very active under very different conditions and difficult to inactivate. At low salt-concentrations (up to 100 mM NaCl), RNase A cleaves single- and double-stranded RNA and RNA in RNA \: DNA- hybrides. Under high salt concentrations (>300 mM NaCl) single-stranded RNA is cleaved only. To remove the enzyme from samples, it has to be digested by proteinase K (frequently, SDS at a final concentration of 0.6 % is added) and several phenol extractions are required. (Applications\: Enzymatic manipulation of DNA and RNA\: ref. 1 Suppl. 8 p. 3.13.1; minipreps of plasmid-DNA\: ref. 1 Suppl. 24 p. 1.6.6; inSitu-hybridisation of cellular RNA\: ref. 1 Suppl. 7 p. 14.3.8; removal of RNA from plasmid preparations\: ref. 2 p. 1.51)Stock solutions are prepared at concentrations from 1 - 10 mg/ml in 10 mM Tris · HCl, pH 7.5; 15 mM NaCl or in 10 mM Tris · HCl, pH 7.5; 1 mM EDTA, pH 8.0 (TE buffer). The recommended working concentration is 10 μg/ml (removal of RNA from plasmid preparations; 1 hr, RT) or 100 ng/ml (preparation of "blunt ends" of double-stranded cDNA).Unit-definition\: One unit of activity is defined as that amount of enzyme which causes the hydrolysis of RNA to yield a velocity constant, k = 1, at 25°C and pH 5.0 (Kunitz-Unit). Inactivation of DNase activity\: A protocol (ref. 2) suggests to dissolve 10 mg/ml RNase A in 0,01 M Sodium acetate (pH 5,2), to heat to 100°C for 15 minutes in a water bath and to cool down to room temperature very slowly. The pH value is equilibrated by adding 0.1-fold the volume of 1 M Tris-Cl (pH 7,4). Caution: Heating solutions of RNase A to inactivate DNase may not be satisfactory since RNase activity may be lost if precipitate formation occurs. For applications that require DNase-free RNase A we recommend our product A3832, RNase A (DNase-free). Stability\: RNase A aggregates during lyophilizing and storage. It has a high affinity to glas surfaces, which has to be taken into consideration. At neutral pH (e. g. in PBS pH 7.4) and high concentrations (> 10 mg/ml) the enzyme will precipitate. At \+4°C (lyophilized) it is stable for several years (dry storage), in solution (-20°C) several years or (\+4°C) several weeks.
Bibliography:
(1)Ausubel, F.A., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. & Struhl, K. (eds.) (1995) Current Protocols in Molecular Biology. Page 3.13.1 Suppl. 8; Greene Publishing & Wiley-Interscience, New York (2)Sambrook, J. & Russel, D.W. (2001) Molecular Cloning\: A Laboratory Manual, 3rd Edition Page A4.39. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. (3)Melton, D.A. et al. (1984) Nucleic Acids Res. 12, 7035-7056Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing the SP6 promoter. (4)Winter, E. et al. (1985) Proc. Natl. Acad. Sci. USA 82, 7575-7579A method to detect and characterize point mutations in transcribed genes.
WGK:
1
Storage:
-20°C
Origin:
from bovine pancreas
EINECS:
232-646-6
CS:
35079090
Download TDS file for complete specifications