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Merck
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70663

Millipore

蛋白酶K,冻干

Highly active serine protease that exhibits broad cleavage specificity on native and denatured proteins and is widely used in the purification of DNA and RNA.

同義詞:

蛋白酶 K 来源于林伯氏白色念球菌, 肽链内切酶

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About This Item

CAS號碼:
酶委員會編號:
MDL號碼:
分類程式碼代碼:
12352202
NACRES:
NA.85

品質等級

形狀

solid

分子量

28.93 kDa

製造商/商標名

Novagen®

儲存條件

OK to freeze

技術

DNA extraction: suitable

運輸包裝

wet ice

儲存溫度

2-8°C

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一般說明

蛋白酶K是从真菌林伯氏白色念球菌中分离出的高活性28,904-Da丝氨酸蛋白酶。该酶对天然和变性蛋白质具有广泛的切割特异性,广泛用于DNA和RNA的纯化。在变性剂如SDS(1%)和高温(50~60°C)存在下,其活性增加。对于蛋白质去除和酶灭活,推荐的工作浓度为50-100 µg/mL,对于组织处理,推荐的工作浓度为2 mg/mL。蛋白酶K冻干粉可制备成20 mg/ml的水溶液,并在-20°C下分装储存。该酶也可作为即用型浓缩储备液(600 mAU/ml),在大多数应用中可方便常规使用。1 mg蛋白酶K相当于30 mAU(AU=Anson单位)。Novagen蛋白酶K产品不含可检测的DNase和RNase

應用

在分离DNA和RNA的过程中,用于核酸酶的蛋白水解失活。
用于去除内毒素结合的阳离子蛋白,如溶菌酶和核糖核酸酶A。
据报道,其可用于分离肝、酵母和绿豆线粒体
用于测定酶在细胞膜上的定位
用于石蜡包埋组织切片的处理,暴露用于抗体标记的抗原结合位点。
用于可传播海绵状脑病(TSE)研究中朊病毒脑组织样本蛋白的消化。

生化/生理作用

蛋白酶K是一种稳定且具有高活性的丝氨酸蛋白酶。晶体和分子结构研究证据表明该酶属于枯草素家族,具有活性位点催化三联体(Asp39-His69-Ser224)。它在一系列广泛的环境中均稳定:pH,缓冲盐,去污剂(SDS)以及温度。 在0.1-0.5% SDS存在的情况下,蛋白酶K可保持活性,在不损害分离DNA完整性的前提下,可以消化DNA制剂中的多种蛋白质和核酸酶。

警告

毒性:有害(C)

單位定義

1 AU(AU=Anson单位)定义为在37°C、pH 7.5下每分钟从酪蛋白中释放1.0 µmol(181 µg)酪氨酸的酶量。

法律資訊

NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany

象形圖

Health hazardExclamation mark

訊號詞

Danger

危險分類

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

標靶器官

Respiratory system

儲存類別代碼

11 - Combustible Solids

水污染物質分類(WGK)

WGK 1

閃點(°F)

Not applicable

閃點(°C)

Not applicable


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Yang Sui et al.
Nucleic acids research, 50(12), 6890-6902 (2022-06-25)
Ribonucleotides can be incorporated into DNA during replication by the replicative DNA polymerases. These aberrant DNA subunits are efficiently recognized and removed by Ribonucleotide Excision Repair, which is initiated by the heterotrimeric enzyme RNase H2. While RNase H2 is essential
Yang Deng et al.
Cell death & disease, 13(12), 1057-1057 (2022-12-21)
Acute ischemic stroke (AIS) is a global cerebrovascular disease with high disability and mortality, which has no effective therapy. Studies have demonstrated that astrocyte-derived exosomes (ADEXs) provided neuroprotection in experimental stroke models. Nevertheless, the role of exosomes derived from oxygen-glucose-deprivation/reoxygenation-stimulated
Jae Wook Hyeon et al.
Scientific reports, 5, 14944-14944 (2015-10-10)
Prion diseases are associated with the conformational conversion of the physiological form of cellular prion protein (PrP(C)) to the pathogenic form, PrP(Sc). Compounds that inhibit this process by blocking conversion to the PrP(Sc) could provide useful anti-prion therapies. However, no
Rodrigo Ibarra-García-Padilla et al.
STAR protocols, 2(3), 100709-100709 (2021-08-18)
Characterizing mRNA and protein expression with temporal and spatial resolution is a valuable component of nearly every developmental study. Here, we describe a protocol that combines in situ hybridization chain reaction (HCR) and immunofluorescence, allowing for the detection of mRNAs and
Felicia Lazure et al.
Nature communications, 14(1), 535-535 (2023-02-03)
Adult stem cells are indispensable for tissue regeneration, but their function declines with age. The niche environment in which the stem cells reside plays a critical role in their function. However, quantification of the niche effect on stem cell function

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