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- 库存:
10
- 英文名:
Proteinase K
- 保质期:
18个月
- 供应商:
嵘崴达
- 保存条件:
2-8度
- 规格:
100MG
位于慕尼黑附近Penzberg的罗氏工厂是全球最大 的生化试剂、诊断试剂和药品的生产基地之一。出产的产品除罗氏自用外,还作为原材料提供给其他的生产厂家。
根据客户的要求,产品可在GMP和GLP标准下生产,并符合FDA,USDA和BGA的规定。厂房通过ISO 9001论证。
一. PCR级重组蛋白酶K
性能优势:1.GMP级别;2.可在室温下长期稳定保存;3.有冻干粉和液态产品供选择。
可根据客户需求提供不同包装的工业原料,价格实惠。
主要通用包装有:03 508 838 103 100 mg(常备现货) 05 963 117 103 5 g
Proteinase K, recombinant, PCR Grade
from Titrirachium album, expressed in Pichia pastoris,
lyophilizate
Application
Proteinase K, recombinant, PCR Grade, digests native proteins very efficiently.
This enzyme can be used to rapidly inactivate endogenous RNases and
DNases during nucleic acid isolation. Proteinase K is particularly suited for the
isolation of native RNA and DNA from tissues and cell lines.
The enzyme promotes cell lysis by activating a bacterial autolytic factor.
Proteinase K is also used for:
■ Analysis of membrane structures by modifying proteins and glycoproteins
on cell surfaces
■ Removal of cellular debris during the preparation of colony lifts
■ Treatment of tissue sections to ensure efficient probe infiltration during in
situ hybridization
Properties
Nomenclature: Proteinase K
Molecular weight: 28.8 kD
pH optimum: 7.5-10.5
Inhibitors: Proteinase K is inhibited by diisopropyl fluorophosphate and phenylmethylsulfonyl
fluoride (PMSF) and is also totally inactivated by mercuric
ions. Pefabloc® SC and Pefabloc® PLUS are specific, irreversible and nontoxic
inhibitors.
Specificity: Proteinase K is one of the most active endopeptidases known and
does not exhibit any pronounced cleavage specificity. Activity can be stimulated
by addition of denaturing agents (SDS and urea).
Specification
Appearance: White lyophilizate
Solubility: Clear, colorless solution in water (c=20 mg/ml)
Volume activity (+37°C, hemoglobin): ≥24 U/mg lyophilizate
Specific activity (+37°C, hemoglobin): ≥30 U/mg protein
Unit definition (hemoglobin): One unit is the enzyme activity which releases
folin positive amino acids and peptides equivalent to 1 μmol of tyrosine in 1
minute under the test conditions.
Volume activity (+25°C, Chromozym): ≥2 U/mg lyophilizate
Specific activity (+25°C, Chromozym): ≥2.5 U/mg protein
Unit definition (Chromozym): One unit is the enzyme activity which cleaves at
+25°C in 1 minute 18 mmol Chromozym TRY.
Unspecific endonucleases (MWM III DNA): Not detectable in up to 200 μg
after 16 hours incubation at +37°C.
Nicking activity (pBR322 DNA): Not detectable in up to 200 μg after 16 hours
incubation at +37°C.
Ribonucleases (MS2 RNA): Not detectable in up to 40 μg after 16 hours
incubation at +37°C.
DNA (Threshold): ≤10 pg/mg enzyme
Bioburden: ≤125 CFU/g
Stability: At +2 to +8°C within specification range for 18 months.
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文献和实验蛋白酶k是一种枯草蛋白酶类的高活性蛋白酶,用于生物样品中蛋白质的一般降解。从林伯氏白色念球菌(tritirachium album limber)中纯化得到。该酶有两个ca2+结合位点,它们离酶的活性中心有一定距离,与催化机理并无直接关系。然而,如果从该酶中除去ca2+,由于出现远程的结构变化,催化活性将丧失80%左右,但其剩余活性通常已足以降解在一般情况下污染酸制品的蛋白质。所以,蛋白酶k消化过程中通常加入edta(以抑制依赖于mg2+的核酸酶的作用
蛋白酶 K,威力巨大的广谱蛋白酶,95C 加热10 分钟,则完全失活。数年前首次使用它,是替代 DEPC 处理 RNA 抽提用的离心管和枪头,效果不错;后来又用于处理 RNase-Free的水,效果也是一级棒。从此就再也不用 DEPC 了。现在将它的细节公开,与大家分享。 配制 RNA 裂解试剂时,直接用灭菌的双蒸水配制,最后,加入蛋白酶 K 至终浓度 1ug/ml。轻轻混匀,室温放置 15 分钟后即可。枪头及离心管去除 RNase:先将枪头及离心管清洗干净后,移入预先混好的含 1ug
编者按: 当我们屏着呼吸使用致癌的DEPC时,是否想过用一种既安全又省钱方法替代DEPC?看过本文,你一定会感慨作者的探索精神。其实,在生物吧,这样的探索还有很多。如果你嫌找得麻烦,那就先看看“牛人强文” 吧! 蛋白酶 K 的妙用,我曾经在别的论坛发表。坦率地说,蛋白酶 K 的这个用途,是我的得意之作之最。不是因为该发现有什么创新,而是因为它的实用性。当别人还在讨论 DEPC










