T1426

Sigma-Aldrich

胰蛋白酶 来源于牛胰腺

TPCK Treated, essentially salt-free, lyophilized powder, ≥10,000 BAEE units/mg protein

CAS号:
EC 号:
MDL编号:
NACRES:
NA.54

质量水平

200

形式

essentially salt-free, lyophilized powder

specific activity

≥10,000 BAEE units/mg protein

分子量

23.8 kDa

溶解性

hydrochloric acid: soluble 1 mM

特色行业

Diagnostic Assay Manufacturing

异质活性

Chymotrypsin ≤0.1 BTEE units/mg protein

储存温度

−20°C

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应用

对于胰蛋白酶消化肽,胰蛋白酶:肽 使用约为 1:100 至 1:20 的比例。从培养表面除去粘附细胞是本产品的一个典型应用。 从其底物中去除细胞所需的胰蛋白酶浓度主要取决于细胞类型和培养物的年龄。 胰蛋白酶也被用于细胞培养期间细胞的再悬浮、蛋白质组学研究中的蛋白质消化和各种凝胶内消化。 其他应用包括:通过基于膜的技术评估结晶,用在确定动力学陷阱的存在会限制蛋白质的折叠速率和产量的一项研究中。

包装

50, 100, 250, 500 mg in glass bottle
1, 5, 100 g in glass bottle

生化/生理作用

胰蛋白酶切割赖氨酸和精氨酸残基的 C 端肽。 如果酸性残基位于切割位点的任一侧,则该反应的水解速度减慢,并且如果脯氨酸残基位于切割位点的羧基侧,则停止水解。 胰蛋白酶活性的最适 pH 为 7-9。 胰蛋白酶还可以起到裂解氨基酸合成衍生物的酯和酰胺键的作用。 将 EDTA 作为螯合剂添加到胰蛋白酶溶液中,用于中和钙离子和镁离子,这些离子遮蔽胰蛋白酶作用的肽键。 去除这些离子会增加酶活性。

丝氨酸蛋白酶抑制剂,包括 DFP、TLCK、APMSF、AEBSEF 和抑肽酶等,将抑制胰蛋白酶。

组分

胰蛋白酶是由223个氨基酸残基的单链多肽所组成,通过从胰蛋白酶原在Lys - lle肽键处被切割去除N末端六肽而产生。 氨基酸的序列是由6个二硫桥所交联而成的。 这是胰蛋白酶、β-胰蛋白酶的天然形式。 β-胰蛋白酶可发生自溶,并在Lys - Ser残基处切割以产生α-胰蛋白酶。 胰蛋白酶是丝氨酸蛋白酶家族的一员。

注意

在1 mM HCl中的溶液经分装并储存在-20°C可稳定1年。Ca2+的存在将也会消除胰蛋白酶的自溶并维持其在溶液中的稳定性。 胰蛋白酶在2.0 M尿素、2.0 M盐酸胍或0.1% (w/v) SDS中可维持其绝大多数的活性。

单位定义

使用 BAEE 作为底物,一个 BAEE 单元,将在 pH 7.6,25℃ 下,每分钟产生 0.001 的 A253

制备说明

可以1 mg/mL溶于1 mM HCl。
它还经TPCK处理并透析。 用L-1-甲苯磺酰胺-2-苯乙基氯甲基酮(TPCK)处理可降低通常存在于胰蛋白酶中的胰凝乳蛋白酶活性。

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 1

个人防护装备

dust mask type N95 (US), Eyeshields, Faceshields, Gloves

分析证书

原产地证书 (CofO)

  1. 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.

  2. 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.

  3. What is Product T1426, Trypsin from bovine pancreas, soluble in, and how do you recommend storing solutions?

    Trypsin is soluble in 1 mM hydrochloric acid (pH approximately 3). It is also surprisingly stable in 1 mM HCl. Solutions are stable for approximately 1 year when aliquoted and stored at -20°C. The presence of Ca2+ (20 mM) will also retard trypsin's ability to autodigest, and will therefore help to maintain the stability of the trypsin in solution.

  4. How much of Product T1426, Trypsin from bovine pancreas, should I use to digest my peptide?

    For trypsin digestion of peptides, use a ratio (w:w) of about 1:100 to 1:20 for trypsin:peptide. Trypsin preparations usually contain some contaminating chymotrypsin and this should be inhibited with N-tosyl-L-phenylalanyl chloromethyl ketone (TPCK). Product No. T1426 has already been treated with TPCK, so it does not need to be further treated.

  5. What is the extinction coefficient for Product T1426, Trypsin from bovine pancreas?

    Various values have been reported in the literature for the extinction coefficient of bovine pancreatic trypsin. The E1%(280) values have ranged from 12.9 - 15.4. When we assay the protein content of Product No. T1426 in our labs here at Sigma, we use a value of 14.4.

  6. Does Product T1426, Trypsin from bovine pancreas, work for in-gel digestions?

    For in-gel digestions, we actually recommend using a methylated trypsin. Product No. T6567 is Trypsin from porcine pancreas, proteomics grade, dimethylated. For product T6567, the lysine residues of proteomics grade trypsin have been reductively methylated, resulting in a product that is resistant to autolysis. The T6567 has also been treated with TPCK to remove chymotryptic activity, further purified through affinity chromatography, and lyophilized. This results in an enzyme that gives highly specific cleavage.

  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.

Jeffrey R Whiteaker et al.
Molecular & cellular proteomics : MCP, 10(4), M110-M110 (2011-01-20)
Stable isotope standards and capture by antipeptide antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution and detection by multiple reaction monitoring mass spectrometry to provide quantitative measurement of peptides as surrogates for their respective proteins. In this...
Yipu Lin et al.
Influenza and other respiratory viruses, 11(3), 263-274 (2017-02-07)
Two new subclades of influenza A(H3N2) viruses became prominent during the 2014-2015 Northern Hemisphere influenza season. The HA glycoproteins of these viruses showed sequence changes previously associated with alterations in receptor-binding properties. To address how these changes influence virus propagation...
Joshua K Endow et al.
PloS one, 11(12), e0167802-e0167802 (2016-12-10)
PolyGly is present in many proteins in various organisms. One example is found in a transmembrane β-barrel protein, translocon at the outer-envelope-membrane of chloroplasts 75 (Toc75). Toc75 requires its N-terminal extension (t75) for proper localization. t75 comprises signals for chloroplast...
Kazuya Sakata et al.
Scientific reports, 6, 37200-37200 (2016-11-16)
The loss-of-function mutations of serine protease inhibitor, Kazal type 1 (SPINK1) gene are associated with human chronic pancreatitis, but the underlying mechanisms remain unknown. We previously reported that mice lacking Spink3, the murine homologue of human SPINK1, die perinatally due...
Philip M Day et al.
The Plant cell, 31(8), 1845-1855 (2019-06-21)
Chloroplasts evolved from a cyanobacterial endosymbiont that resided within a eukaryotic cell. Due to their prokaryotic heritage, chloroplast outer membranes contain transmembrane β-barrel proteins. While most chloroplast proteins use N-terminal transit peptides to enter the chloroplasts through the translocons at...
实验方案
This procedure is for products with a specification for Trypsin activity using Na-Benzoyl-L-arginine ethyl ester (BAEE) as a substrate. The procedure is a continuous spectrophotometric rate determination (A253, Light path = 1 cm).
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相关内容
本方案适用于使用Nα-苯甲酰基-L-精氨酸乙酯(BAEE)作为底物测定胰蛋白酶活性。本方案是基于连续分光光度速率测定(A253,光程= 1cm)方案:
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