P5147

Sigma-Aldrich

蛋白酶 来源于灰色链霉菌

Type XIV, ≥3.5 units/mg solid, powder

别名:
Actinase E, Pronase E
CAS号:
EC 号:
MDL编号:
eCl@ss:
32160410

质量水平

200

类型

Type XIV

形式

powder

specific activity

≥3.5 units/mg solid

溶解性

10 mM NaAc (pH 7.5) and 5 mM CaAc: soluble 0.2 mg/mL at 37 °C, clear, colorless to tan

运输

wet ice

储存温度

−20°C

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相关类别

特异性

一种具有包括细胞外丝氨酸蛋白酶在内至少三种蛋白水解活性的混合物。通常,丝氨酸蛋白酶可显示出广泛的底物特异性,而这被视为通过分子中由一个Asp、一个His和Ser残基组成的活性位点所介导的。该酶更倾向于水解谷氨酸或天冬氨酸的羧基侧的肽键。

应用

蛋白酶是一种通过水解肽键来分解蛋白的酶。 蛋白酶用于降解蛋白、研究蛋白酶抑制剂和研究热灭活动力学。 蛋白酶用于孵育中核酸的分离过程。 来自灰色链霉菌的蛋白酶已用于通过四肽醛和丝氨酸蛋白酶而形成的共价复合物的结晶学和动力学研究中。
来自灰色链霉菌的蛋白酶已被用于:
  • 消化髓核(NP)组织
  • 催化不对称曼尼希一锅反应
  • 淀粉脱蛋白
  • 消化细胞解离用脑切片

蛋白酶通常用于核酸的分离过程,通过补充0.2%十二烷基硫酸钠和10 mM EDTA并孵育0.5-3.0小时。
该酶比已知碱性蛋白酶在更高的pH范围内更具活性,即使在0.2N NaOH溶液中也显示出了蛋白水解活性。该酶用于不溶性蛋白的蛋白水解和蛋白的结构研究。
来自Sigma的酶已用于抗凝血酶-肝素复合物的消化和分析。它还用于分离抗酶淀粉。

包装

100 mg in glass bottle
1, 5 g in glass bottle

生化/生理作用

蛋白酶可通过肽键的水解使蛋白质分解代谢。 蛋白酶可被丝氨酸活性位点抑制剂所灭活,如苯甲基磺酰氟(PMSF)和二异丙基氟磷酸盐。 来自灰色链霉菌的蛋白酶是一种具有包括细胞外丝氨酸蛋白酶在内至少三种蛋白水解活性的混合物。丝氨酸蛋白酶显示出广泛的底物特异性,而这被视为通过分子中由一个Asp、一个His和Ser残基组成的活性位点所介导的。该酶更倾向于水解谷氨酸或天冬氨酸的羧基侧的肽键。
该产品是由至少三种酪蛋白水解活性和一种氨肽酶活性所组成的混合物。该酪蛋白分解酶被命名为灰色链霉菌蛋白酶A、灰色链霉菌蛋白酶B和灰色链霉菌胰蛋白酶。当需要广泛或完全降解蛋白质时,可使用该产品。该蛋白酶混合物是高度非特异性的并可作为单氨基酸消化酪蛋白至>70%的程度。

特点和优势

  • 在pH 5.0中9.0范围内高度稳定,在pH 8.8具有最高活性
  • 与多种DNA和RNA分离缓冲液兼容
  • 广泛的底物特异性

质量

含有醋酸钙。

物理属性

通过在80 °C以上加热15-20分钟以完全灭活。

单位定义

在pH 7.5,37 °C 条件下,一单位的酶每分钟将会水解酪蛋白以产生与1.0 μ摩尔 (181 μg) 酪氨酸等同的颜色(Folin-Ciocalteu试剂提供的颜色)。

制备说明

收集自灰色链霉菌的培养基肉汤中。

象形图

Exclamation markHealth hazard

警示用语:

Danger

hazcat

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

靶器官

Respiratory system

storage_class_code

11 - Combustible Solids

WGK Germany

WGK 2

闪点(F)

Not applicable

闪点(C)

Not applicable

个人防护装备

Eyeshields, Gloves, type N95 (US)

分析证书

原产地证书 (CofO)

  1. Is Product P5147, Protease from Streptomyces griseus, also described as "Pronase"?

    "Pronase E" is the name given to a group of proteolytic enzymes produced by Streptomyces griseus K-1. At least 10 proteases are in the mixture, five serine-type proteases, two zinc endopeptidases, two zinc leucine aminopeptidases, and one zinc carboxypeptidase.

  2. Does Product P5147, Protease from Streptomyces griseus, contain calcium acetate?

    This product contains calcium acetate buffer salt. The amount of calcium acetate present will vary for each lot number and has been observed to range from 2 - 30% (w/w).

  3. How should Product P5147, Protease from Streptomyces griseus, be reconstituted and how should a stock solution be prepared?

    This product can be dissolved in 0.01 M sodium acetate with 0.005 M calcium acetate at pH 7.5 at 37°C; at 0.2 mg/mL, this yields a clear solution which ranges from colorless to light tan. Calcium ion is recommended for protection from autolysis. The activity of a dilute enzyme solution containing 0.01 to 0.1 M calcium ion was stable over 24 hours at neutral pH at 2-8°C.  Pronase E is stable at 4°C for at least six months.  Stock solutions of 5 to 20 mg/mL in water are usually stored at -20°C.

  4. Under what conditions can Product P5147, Protease from Streptomyces griseus, be used to digest protein?

    For DNA isolation, Pronase E is usually prepared as a stock solution, and prior to storage at -20°C, the solution is first heated to 56°C for about 15 minutes, then incubated at 37°C for 1 hour. This encourages self-digestion, to eliminate DNAse and RNAse contamination. The enzyme is added to a DNA sample (in the presence of 0.5-1%(w/v) SDS to disrupt DNA protein interactions) typically at 250-500 μg protein/mL, then incubated at 37°C for 1-4 hours. For protein hydrolysis, dissolve about 0.2 μmol of protein in 0.2 mL of 50 mM ammonium bicarbonate buffer at pH 8 (or phosphate buffer pH 7). Add Pronase to 1% (w/w) and incubate at 37°C for 24 hours. It may be necessary to add aminopeptidase M at 4% (w/w) and incubate at 37°C for another 18 hours.

  5. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  6. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  7. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  8. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  9. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

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X-ray crystallographic data show that a specific tetrapeptide aldehyde inhibitor (N-acetylprolylalanylprolylphenylalaninal) forms a stable, covalent, tetrahedral addition complex with the serine protease, SGPA, from Streptomyces griseus. Earlier proposals, based on kinetic measurements, for the covalent nature of such linkages are...
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Xue Y, et al.
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实验方案
To standardize a procedure for the enzymatic assay of Protease using Casein as a substrate.
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相关内容
建立以酪蛋白为底物的蛋白酶的标准酶法测定方法。
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