产品名称 : | PH0624 | RNase A 核糖核酸酶A |
产品品牌 : | 飞净 PHYGENE |
将RNase A粉末溶解在0.01M NaAc PH5.2中,使终浓度为10mg/mL,于100℃沸水浴中加热15min,室温下缓慢冷却 。用0.1体积的1M Tris-HCl 调PH值至7.4左右,分装-20℃保存。当浓缩的RNase A溶液在中性PH条件下加热到100℃时,RNase A 产生沉淀。(注:引自分子克隆实验指南第三版 1647页)
Product Description
RNase A is an endoribonuclease that attacks at the 3’ phosphate of a pyrimidine nucleotide. The sequence of pG-pG-pC-pA-pG will be cleaved to give pG-pG-pCp and A-pG. The highest activity is exhibited with single stranded RNA. RNase A is a single chain polypeptide containing 4 disulfide bridges. In contrast to RNase B, it is not a glycoprotein.
RNase A can be inhibited by alkylation of His12 or His119, which are present in the active site of the enzyme. Activators of RNase A include potassium and sodium salts. Molecular mass:~13.7 kDa (amino acid sequence) Isoelectric point: pI = 9.6
Preparation
An RNase A solution can be made free of DNase by boiling. According to a literature method, prepare a 10 mg/mL stock solution in 10 mM sodium acetate buffer, pH 5.2. Heat to 100℃ for 15 minutes, allow to cool to room temperature, and then adjust to pH 7.4 using 0.1 volume of 1 M Tris-HCl, pH 7. 4. Aliquot and store at –20 ℃. If RNase A is boiled at a neutral pH, precipitation will occur. When boiled at the lower pH, some precipitation may occur because of protein impurities that are present.
Stability
Store at Rnase A at –20 ℃. Stock solutions stored in frozen aliquots remain active for at least 6 months. RNase A is a very stable enzyme and solutions have been repo rted to withstand temperatures up to 100 ℃. At 100 ℃, an RNase A solution is most stable between pH 2.0 and 4.5.
Procedure
A major application for RNase A is the removal of RNA from preparations of plasmid DNA. For this application, DNase free RNase A is used at a final concentration of 10μg/mL.
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