Wako和光纯药 125-05061 赖氨酰肽链内切酶
货号：125-05061
包装：20ug x5
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。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
|
|
佳pH
|
9.0-9.5（酰胺酶的佳活性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
- Wada, Y., and Kadoya, M.: J. Mass Spectrom., 38, 117 (2003).
- 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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145-08221 Nuclease P1 for Genetic Research 500u 54576-84-0
162-09115 Polyethylene Glycol 4,000 Wako 1st Grade 500g 25322-68-3
164-21655 N/400 Potassium Polyvinyl Sulfate Solution for Colloidal Titration 500mL 26837-42-3
165-09105 Polyethylene Glycol 2,000 Wako 1st Grade 500g 25322-68-3
165-17035 Polyvinylpyrrolidone K 30 Wako Special Grade 500g 9003-39-8
167-25501 PA tag Peptide for Genetic Research 5mg
169-04245 Potassium Dihydrogen Phosphate JIS Special Grade 500g 7778-77-0
169-09125 Polyethylene Glycol 6,000 Wako 1st Grade 500g 25322-68-3
169-27261 PA tag Washing Solution for Genetic Research 50mL
185-01003 Rifampicin for Biochemistry 5g 13292-46-1
186-01114 all-trans-Retinoic Acid for Biochemistry 50mg 302-79-4
189-01001 Rifampicin for Biochemistry 1g 13292-46-1
190-05535 Trisodium Citrate Dihydrate for Amino Acid Automated Analysis 500g 6132-4-3
191-01665 Sodium Chloride JIS Special Grade 500g 7647-14-5
191-02885 Trisodium Phosphate 12-Water JIS Special Grade 500g 10101-89-0
193-02041 Glycerol 2-Phosphate Disodium Salt n-Hydrate 100g 819-83-0
195-12854 Streptavidin, from Streptomyces avidinii for Immunochemistry 5mg 9013-20-1
195-16031 StemSure Freezing Medium for Cell Culture 100mL
196-00015 Sucrose JIS Special Grade 500g 57-50-1
196-08675 Sodium Dodecyl Sulfate Wako 1st Grade 500g 151-21-3
197-07485 Sodium Sulfate for PCB and Phthalic Acid Esters Analysis 500g 7757-82-6
199-02825 Sodium Dihydrogenphosphate Dihydrate Wako 1st Grade 500g 13472-35-0
203-17561 Trichostatin A for Cellbiology 1mg 58880-19-6
207-00055 L(+)-Tartaric Acid JIS Special Grade 500g 87-69-4
208-08385 1mol/L Triethylammonium Hydrogencarbonate Solution for Nucleic Acid Synthesis 500mL 15715-58-9
223-02112 VA-044 for Cellbiology 25g 27776-21-2
225-02111 VA-044 for Cellbiology 100g 27776-21-2
227-01071 Valproic Acid for Biochemistry 5g 99-66-1
230-01981 Wetting Tension Test Mixture No.46.0 for Determination of Wetting Tension 50mL
231-01911 Wetting Tension Test Mixture No.39.0 for Determination of Wetting Tension 50mL