Wako和光纯药 125-05061 赖氨酰肽链内切酶

Wako和光纯药 125-05061 赖氨酰肽链内切酶

货号:125-05061
包装:20ug x5

  • 英文名:Lysyl Endopeptidase, Mass Spectrometry Grade
  • 级别:for Proteome Research
  • 分子量:27,500
  • 品牌:Wako
  • 外观:Lyophilisate. Keep at -20 degrees C.

赖氨酰肽链内切酶(Lysyl Endopeptidase)可以特异性剪切除赖氨酸残基C末端和S-氨乙基半胱氨酸残基的肽键,用于蛋白测序和Lys-X 化合物的酶催化合成。该酶(#125-05061)稳定性高,在4M 尿素或0.1%SDS 溶液中30度孵育6小时之后,仍然拥有完整的生物活性。

*上海金畔生物提示:我司所销售的化学试剂、原料等所有产品(包括但不限于抗生素类、蛋白质类、试剂盒类产品等)仅限用于科学研究用途,不得作用于人体。

质谱级 赖氨酰肽链内切酶 Lysyl Endopeptidase
赖氨酰肽链内切酶(Lysyl Endopeptidase)最初由Masaki等人从土壤细菌中分离得到。该酶可以特异性剪切除赖氨酸残基C末端和S-氨乙基半胱氨酸残基的肽键,用于蛋白测序和Lys-X 化合物的酶催化合成。作为Wako官方授权分销商,上海金畔生物专业提供Wako多种级别赖氨酰肽链内切酶,该酶稳定性高,在4M 尿素或0.1%SDS 溶液中30℃孵育6小时之后,仍然拥有完整的生物活性。
质谱级赖氨酰肽链内切酶是质谱分析前处理时最常用的蛋白分解酶。若同时使用赖氨酰肽链内切酶和胰酶,可更好地切断赖氨酸基团的多肽,增加多肽的数量。产品已按照使用习惯做成小包装,是方便使用的冷冻干燥品。
质谱级 赖氨酰肽链内切酶 Lysyl Endopeptidase 理化特性
表1赖氨酰肽链内切酶理化特性表
外观
冻干粉(包含2 mmol/L Tris-HCl buffer,pH 8)
活性
0.03~0.07AU/vial
分子量
27,000(凝胶过滤);30,000 (SDS-PAGE)
溶解性
易溶于水或缓冲液
最佳pH
9.0-9.5(酰胺酶的最佳活性pH)
等电点
6.9-7.0
抑制剂
DFP、PMSF、TLCK
来源
细菌
稳定性
溶于pH 值5.0-12.0 的Tris缓冲液中,可于4℃稳定保存2年。溶于pH 值6.0-11.0 的缓冲液中,可于30℃稳定保存,但是50℃及以上不稳定。
单位定义
一单位酰胺酶(AU) 指在30℃ pH 9.5 时每分钟产生1 μmol 对硝基苯胺所需的酶量。
底物特异性
水解底物:Tos-Lys-Ome、Bz-Lys-NH2、Bz-Lys-pNA、Lys-pNA
非水解底物:Bz-Arg-NH2、Bz-Arg-pNa、Arg-pNA
质谱级 赖氨酰肽链内切酶 Lysyl Endopeptidase 产品特点
  • 高特异性、高蛋白质消化率、适用于蛋白质谱分析
  • 提高裂解效率、增加肽段数量
  • 根据使用量特意制备小包装,方便使用
质谱级 赖氨酰肽链内切酶 Lysyl Endopeptidase 应用实例
分别采用胰蛋白酶(Tp)、赖氨酰肽链内切酶(Lep) 和Tp 与Lep联用进行胶内酶切的效果比较:牛血清蛋白BSA 的条带(100ng) 通过SDS-PAGE 获得,然后分别用Tp、Lep 和Lep+Tp 进行酶切,再用MALDI-TOFMS 法进行分析。蛋白酶的实验效果如下表:
表2 Tp、Lep 和 Lep+Tp 的蛋白酶酶切结果对照
Tp
Lep
Lep +Tp
裂解位点
精氨酸和赖氨酸的C端
赖氨酸的C端
精氨酸和赖氨酸的C端
错误裂解率
(错误裂解所占比例)
多(8%)
很少(0%)
少(3%)
鉴定出的多肽数量
17
19
22
结果表明 Lep 酶解的错误裂解率最低,Tp酶解时加入Lep后,错误裂解率有所降低,同时,可以鉴定出更多的多肽。

图1 胰蛋白酶 (Tp)(图 a) 和赖氨酰肽链内切酶 (Lep)+Tp (图 b) 酶切后的质谱结果对照图。

Lep+Tp酶切后,可以在m/z=2000时得到吸收峰,而单独的Tp酶切在m/z=2000时没有吸收峰。该结果表明Lep可以提高测序覆盖度。
( 数据由大阪医疗中心和妇婴健康研究所Y. Wada博士提供 )
质谱级 赖氨酰肽链内切酶 Lysyl Endopeptidase 产品列表
品牌
产品编号
产品名称
产品规格
产品等级
Wako
125-05061
Lysyl Endopeptidase, MS Grade
赖氨酰肽链内切酶,MS级
20μgx5
质谱级
Wako
125-02543
Lysyl Endopeptidase (R)
赖氨酰肽链内切酶
1vial(2AU)
for Biochemistry
Wako
129-02541
Lysyl Endopeptidase (R)
赖氨酰肽链内切酶
1vial(10AU)
for Biochemistry
质谱级 赖氨酰肽链内切酶 Lysyl Endopeptidase 参考文献
  1. Wada, Y., and Kadoya, M.: J. Mass Spectrom., 38, 117 (2003).
  2. Shevchenko, A., Wilm, MM., Vorm, O., and Mann., M.: Anal. Chem., 68, 850 (1996).
上海金畔生物专业提供Wako赖氨酰肽链内切酶,正品保障,值得您信赖!
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