Wako和光純藥 125-05061 賴氨酰肽鏈內(nèi)切酶
貨號(hào)：125-05061
包裝：20ug x5
包裝：20ug x5
- 英文名：Lysyl Endopeptidase， Mass Spectrometry Grade
- 級(jí)別：for Proteome Research
- 分子量：27，500
- 品牌：Wako
- 外觀：Lyophilisate. Keep at -20 degrees C.
賴氨酰肽鏈內(nèi)切酶（Lysyl Endopeptidase）可以特異性剪切除賴氨酸殘基C末端和S-氨乙基半胱氨酸殘基的肽鍵，用于蛋白測(cè)序和Lys-X 化合物的酶催化合成。該酶（#125-05061）穩(wěn)定性高，在4M 尿素或0.1%SDS 溶液中30度孵育6小時(shí)之后，仍然擁有完整的生物活性。
*上海金畔生物提示：我司所銷售的化學(xué)試劑、原料等所有產(chǎn)品（包括但不限于抗生素類、蛋白質(zhì)類、試劑盒類產(chǎn)品等）僅限用于科學(xué)研究用途，不得作用于人體。
質(zhì)譜級(jí) 賴氨酰肽鏈內(nèi)切酶 Lysyl Endopeptidase
賴氨酰肽鏈內(nèi)切酶（Lysyl Endopeptidase）最初由Masaki等人從土壤細(xì)菌中分離得到。該酶可以特異性剪切除賴氨酸殘基C末端和S-氨乙基半胱氨酸殘基的肽鍵，用于蛋白測(cè)序和Lys-X 化合物的酶催化合成。上海金畔生物專業(yè)提供Wako多種級(jí)別賴氨酰肽鏈內(nèi)切酶，該酶穩(wěn)定性高，在4M 尿素或0.1%SDS 溶液中30℃孵育6小時(shí)之后，仍然擁有完整的生物活性。
質(zhì)譜級(jí)賴氨酰肽鏈內(nèi)切酶是質(zhì)譜分析前處理時(shí)最常用的蛋白分解酶。若同時(shí)使用賴氨酰肽鏈內(nèi)切酶和胰酶，可更好地切斷賴氨酸基團(tuán)的多肽，增加多肽的數(shù)量。產(chǎn)品已按照使用習(xí)慣做成小包裝，是方便使用的冷凍干燥品。
質(zhì)譜級(jí) 賴氨酰肽鏈內(nèi)切酶 Lysyl Endopeptidase 理化特性
表1賴氨酰肽鏈內(nèi)切酶理化特性表
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外觀
|
凍干粉（包含2 mmol/L Tris-HCl buffer，pH 8）
|
活性
|
0.03~0.07AU/vial
|
|
分子量
|
27,000（凝膠過(guò)濾）；30,000 (SDS-PAGE)
|
溶解性
|
易溶于水或緩沖液
|
|
佳pH
|
9.0-9.5（酰胺酶的佳活性pH）
|
等電點(diǎn)
|
6.9-7.0
|
|
抑制劑
|
DFP、PMSF、TLCK
|
來(lái)源
|
細(xì)菌
|
|
穩(wěn)定性
|
溶于pH 值5.0-12.0 的Tris緩沖液中，可于4℃穩(wěn)定保存2年。溶于pH 值6.0-11.0 的緩沖液中，可于30℃穩(wěn)定保存，但是50℃及以上不穩(wěn)定。
|
||
|
單位定義
|
一單位酰胺酶(AU) 指在30℃ pH 9.5 時(shí)每分鐘產(chǎn)生1 μmol 對(duì)硝基苯胺所需的酶量。
|
||
|
底物特異性
|
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
|
|||
質(zhì)譜級(jí) 賴氨酰肽鏈內(nèi)切酶 Lysyl Endopeptidase 產(chǎn)品特點(diǎn)
- 高特異性、高蛋白質(zhì)消化率、適用于蛋白質(zhì)譜分析
- 提高裂解效率、增加肽段數(shù)量
- 根據(jù)使用量特意制備小包裝，方便使用
質(zhì)譜級(jí) 賴氨酰肽鏈內(nèi)切酶 Lysyl Endopeptidase 應(yīng)用實(shí)例
分別采用胰蛋白酶(Tp)、賴氨酰肽鏈內(nèi)切酶(Lep) 和Tp 與Lep聯(lián)用進(jìn)行膠內(nèi)酶切的效果比較：牛血清蛋白BSA 的條帶(100ng) 通過(guò)SDS-PAGE 獲得，然后分別用Tp、Lep 和Lep+Tp 進(jìn)行酶切，再用MALDI-TOFMS 法進(jìn)行分析。蛋白酶的實(shí)驗(yàn)效果如下表：
表2 Tp、Lep 和 Lep+Tp 的蛋白酶酶切結(jié)果對(duì)照
|
Tp
|
Lep
|
Lep +Tp
|
|
|
裂解位點(diǎn)
|
精氨酸和賴氨酸的C端
|
賴氨酸的C端
|
精氨酸和賴氨酸的C端
|
|
錯(cuò)誤裂解率
(錯(cuò)誤裂解所占比例) |
多(8%)
|
很少(0%)
|
少(3%)
|
|
鑒定出的多肽數(shù)量
|
17
|
19
|
22
|
結(jié)果表明 Lep 酶解的錯(cuò)誤裂解率最低，Tp酶解時(shí)加入Lep后，錯(cuò)誤裂解率有所降低，同時(shí)，可以鑒定出更多的多肽。
圖1 胰蛋白酶 (Tp)(圖 a) 和賴氨酰肽鏈內(nèi)切酶 (Lep)+Tp (圖 b) 酶切后的質(zhì)譜結(jié)果對(duì)照?qǐng)D。
Lep+Tp酶切后，可以在m/z=2000時(shí)得到吸收峰，而單獨(dú)的Tp酶切在m/z=2000時(shí)沒(méi)有吸收峰。該結(jié)果表明Lep可以提高測(cè)序覆蓋度。
( 數(shù)據(jù)由大阪醫(yī)療中心和婦嬰健康研究所Y. Wada博士提供 )
質(zhì)譜級(jí) 賴氨酰肽鏈內(nèi)切酶 Lysyl Endopeptidase 產(chǎn)品列表
|
品牌
|
產(chǎn)品編號(hào)
|
產(chǎn)品名稱
|
產(chǎn)品規(guī)格
|
產(chǎn)品等級(jí)
|
|
Wako
|
125-05061
|
Lysyl Endopeptidase, MS Grade
賴氨酰肽鏈內(nèi)切酶，MS級(jí) |
20μgx5
|
質(zhì)譜級(jí)
|
|
Wako
|
125-02543
|
Lysyl Endopeptidase (R)
賴氨酰肽鏈內(nèi)切酶 |
1vial(2AU)
|
for Biochemistry
|
|
Wako
|
129-02541
|
Lysyl Endopeptidase (R)
賴氨酰肽鏈內(nèi)切酶 |
1vial(10AU)
|
for Biochemistry
|
質(zhì)譜級(jí) 賴氨酰肽鏈內(nèi)切酶 Lysyl Endopeptidase 參考文獻(xiàn)
- 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).
上海金畔生物專業(yè)提供Wako賴氨酰肽鏈內(nèi)切酶，正品保障，值得您信賴！
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078-05961 G-418 Sulfate for Genetic Research 1g 108321-42-2
093-06471 Insulin, Human, recombinant 50mg
097-06474 Insulin, Human, recombinant for Cell Culture 1g 11061-68-0
120-04891 L-012 for Biochemistry 100mg
125-02543 Lysyl Endopeptidase for Biochemistry 1vial(2AU)
125-05061 Lysyl Endopeptidase, Mass Spectrometry Grade for Proteome Research 5×20μg –
127-06621 Lysyl Endopeptidase Solution for Biochemistry 1EA(5AU) 123175-82-6
129-02541 Lysyl Endopeptidase for Biochemistry 10AU
129-04861 LY 294002 for Biochemistry 5mg 154447-36-6
130-07871 Melibiose for Biochemistry 5g 585-99-9
137-06085 Molecular Sieves 4A1/16 ?500g 70955-01-0
138-07872 Melibiose for Biochemistry 25g 585-99-9
238-01921 Wetting Tension Test Mixture No.40.0 for Determination of Wetting Tension 50mL
238-02021 Wetting Tension Test Mixture No.54.0 for Determination of Wetting Tension 50mL
238-02141 Wetting Tension Test Mixture No.73.0 for Determination of Wetting Tension 50mL 7732-18-5
253-00513 Y-27632 for Cellbiology 5mg 331752-47-7
257-00511 Y-27632 for Cellbiology 1mg 331752-47-7
257-00631 YM-254890 for Cellbiology 1mg 568580-02-9
290-34251 Presep-C DNPH for Sample Pretreatment 20EA
290-63701 LabAssay Triglyceride for Cellbiology 1000tests
290-65901 LabAssay Creatinine for Cellbiology 500tests
291-58601 Lab Assay ALP for Cellbiology 900tests
293-51601 Endotoxin Extracting Solution for LAL Test for Endotoxin Detection 10mL×4 –
293-73201 ScreenFectA for Genetic Research 0.2ml
293-77101 ScreenFect A plus for Genetic Research 0.2ml
293-77601 MagCapture Exosome Isolation Kit PS for Genetic Research 10tests
294-63601 LabAssay NEFA for Cellbiology 750tests
294-65801 LabAssay Cholesterol for Cellbiology 1000tests
295-50201 DNA Extractor Kit for Genetic Research 50tests
295-51301 Limulus ES-II Single Test Wako for Endotoxin Detection 25tests
295-73401 Fructooligosaccharides Set for High Performance Liquid Chromatography Set
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