SAE0151

プロテイナーゼ K from Tritirachium album

free of DNA contaminants, suitable for Microbiome research, lyophilized powder, ≥30 units/mg protein

• 別名: エンドペプチダーゼK
• CAS番号: 39450-01-6
• Enzyme Commission number: 3.4.21.64 (BRENDA, IUBMB)
• MDL番号: MFCD00132129
• UNSPSCコード: 12352202
• NACRES: NA.77

特性

• 品質水準: 200
• 比活性: 30 units/mg protein
• 分子量: 28.93 kDa
• 特徴: DNA free
• 輸送温度: wet ice
• 保管温度: −20°C

詳細

詳細

Purified Proteinase K DNA free SAE0151 undergoes strict quality control testing to ensure the absence of detectable levels of contaminating DNA using 35 cycles PCR amplification of 16S and 18S rDNA using universal primer sets.

Proteinase K is a stable serine protease with broad substrate specificity. It degrades many proteins in the native state even in the presence of detergents. Proteinase K was isolated from a fungus able to grow on keratin and the enzyme can digest native keratin. The enzyme belongs to the subtilisin family with an active site catalytic triad (Asp39 -His69 -Ser224).
The predominant site of cleavage is the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked alpha amino groups. It is commonly used for its broad specificity. Proteinase K is usually denatured by subsequent phenol extractions.

The study of microbial communities has been revolutionized in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. Since DNA contamination during sample preparation is a major problem of these sequence-based approaches, DNA extraction reagents free of DNA contaminates are essential.

アプリケーション

DNAおよびRNAの分離中にヌクレアーゼのタンパク質分解性不活性化を行うために有用です。

DNAおよびRNA分離中にヌクレアーゼのタンパク質分解性不活性化を行うために有用です。
リゾチーム、リボヌクレアーゼAなどの陽イオン性タンパク質に結合するエンドトキシンを除去します。
肝臓、酵母、リョクトウのミトコンドリアを分離するために有用であることが報告されています
膜上の酵素局在化を評価します
抗体標識のためパラフィン包埋組織切片処理に使用され、抗原結合部位を露出させます。
伝達性海綿状脳症（TSE）研究において、プリオンを調べるために、脳組織サンプルのタンパク質を分解する目的で使用されます。

• Proteinase K is frequently used in molecular biology applications to digest unwanted proteins, such as nucleases from DNA or RNA preparations from microorganisms, cultured cells, and plants.
• Removes endotoxins that bind to cationic proteins such as lysozyme and ribonuclease A useful for the isolation of hepatic, yeast, and mung bean mitochondria.
• Determination of enzyme localization on membranes
• Treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling.
• Digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research.

生物化学的／生理学的作用

プロテイナーゼＫは安定した反応性の高いセリンプロテアーゼです。結晶構造および分子構造の研究で得られたエビデンスは、この酵素がスブチリシンファミリーに属し、活性部位が触媒三残基造（Asp³⁹-His⁶⁹-Ser²²⁴）を持つことを示しています。プロテイナーゼＫは広範囲の環境（pH、バッファー塩、界面活性剤［SDS］、温度）で安定です。 0.1～0.5%のSDSの存在下でプロテイナーゼKは活性を保ち、DNA調製において分離されたDNAの完全性を損なうことなくさまざまなタンパク質およびヌクレアーゼを分解します。

特徴および利点

Purified Proteinase K DNA free SAE0151 undergoes strict quality control testing to ensure it will be free of DNA contaminants, suitable for Microbiome research.

単位の定義

One unit will hydrolyze urea denatured hemoglobin to produce color equivalent to 1.0 micromole of tyrosine per minute at pH 7.5, 37°C (color per Folin-Ciocalteu reagent).

調製ノート

Proteinase K is active in 1% Triton X-100 and in 0.5% (w/v) SDS. SDS and urea will denature protein substrates resulting in increased digestion rates. Proteinase K itself is denatured much more slowly by these agents.

安全性情報

• ピクトグラム: GHS08,GHS07
• シグナルワード: Danger
• 危険有害性情報: H315,H319,H334,H335
• 注意書き: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• 危険有害性の分類: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• ターゲットの組織: Respiratory system
• 保管分類コード: 11 - Combustible Solids
• WGK: WGK 1
• 引火点(°F): Not applicable
• 引火点(℃): Not applicable