蛋白酶－C 产品/ Proteases-C Products
Trypsin 蛋白酶, 质谱级
Trypsin Protease,MS Grade
订货号：HLS TRY001C
说明: 胰蛋白酶特异性水解在赖氨酸和精氨酸残留物的碳端的肽键。未修饰的胰蛋白酶受制于蛋白酶的自水解的，产生的残余物影响HPLC肽段分析,另外，自水解可以导致假性胰蛋白酶的产物, 显示出胰凝乳蛋白酶的特性。华利世质谱级胰蛋白酶是修饰猪的胰蛋白酶，通过还原甲基化补偿它阻止蛋白水解消化。质谱级修饰胰蛋白酶通过TPCK处理，亲和纯化，进一步改进，生成高活性和稳定的分子
物理形态: 质谱级Trypsin蛋白酶是冻干粉
分子量: 23kDa.
溶解液（HLS HCL001C）：Trypsin蛋白酶复溶解缓冲液建议1mM盐酸
储存条件：冻干粉末存储在-20℃冰箱，再溶解的酶储存在-20℃。有效期见产品标签
保质期：12个月，在-20℃条件下
pH 值范围: pH7-9范围有最大的活性，在pH4翻转无活性
适用范围：Trypsin蛋白酶特异性水解变性后蛋白质K和R位，推荐和rLys-C混合使用。
推荐SOP: 通用溶液酶解法
Trypsin蛋白酶按照 25至100:1 蛋白和蛋白酶的量(w/w)使用, 37°C恒温过夜5-12小时.
温馨提示：
1.为了保证最大活性，使用提供的胰蛋白酶溶解液溶解质谱级胰蛋白酶，使用前在30℃保温15分钟2.尿素的浓度最好小于1M
Part No. Name Size/pkg
HLS TRY001C Trypsin, Mass Spec Grade 100 μg
HLS HCL001 1 mM hydrochloric acid buffer 0.5 mL
Description: Trypsin specifically hydrolyzes peptide bonds at the carboxyl side of lysine and arginine residues. Unmodified trypsin is subject to auto-proteolysis, generating fragments that can interfere with protein sequencing or HPLC peptide analysis. In addition, autoproteolysis can result in the generation of pseudo trypsin, which has been shown to exhibit chymotrypsin-like specificity. Mass Spec Grade modified trypsin is porcine trypsin modified by reductive methylation, rendering it resistant to proteolysis digestion. In enzymatic stability tests, modified trypsin was not found to self-hydrolyze but can retain higher the activity of general trypsin.
Physical Appearance: Lyophilized powder.
Molecular Weight: 23 kDa
Resuspension Buffer (HLS CHL001C): Trypsin Resuspension Buffer is composed of 1mM hydrochloric acid.
Storage Conditions: Store the lyophilized powder at –20 °C; store reconstituted enzyme at –80 °C.
Shelf life: 24 months at -20 ℃
Stability: Modified trypsin is maximally active in the pH range of 7–9 and reversibly inactivated at pH 4.
Usage Notes:
-
For maximum activity, resuspend trypsin in the resuspension buffer provided, and heat at 30 °C for 15 min before use.
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Thaw the reconstituted trypsin at room temperature, placing on ice immediately after thawing.
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Add trypsin to a protease : protein ratio to be in range of 1:100 - 1:25 (w/w) for protein identification (recommended). Mix well and incubated for 37 °C for 4 h.
Activity: 13,397 units/mg
rLys C蛋白酶, 质谱级
rLys C Protease, MS Grade
订货号：HLS LYS001C
说明: 质谱级rLys-蛋白酶特异性水解在赖氨酸片段的C端的肽键。可用于提高对不易水解的蛋白质的消化; 推荐在6-8M尿素溶液中进行
物理形态: 质谱级rLys-C蛋白酶是冻干粉，含0.5mg海藻糖/瓶
分子量: 27kDa.
溶解液（HLS HAc001C）：rLys-C蛋白酶复溶解缓冲液建议1mM盐酸或50mM醋酸
储存条件：冻干粉末存储在-20℃冰箱，再溶解的酶储存在-20℃。有效期见产品标签
保质期：12个月，在-20℃条件下
pH 值范围: 在PH 8-9 时， rLys-C 具有最佳活性
适用范围：rLys-蛋白酶特异性水解变性后比较不易酶切的蛋白质K位，推荐和Trypsin混合使用。混合酶推荐SOP:
A. 通用溶液酶解法
100ug Trypsin/20ug rLys-C 混合酶按照 25:1 蛋白和蛋白酶的量(w/w)使用, 37°C恒温过夜13-16小时.
温馨提示：尿素的浓度最好小于1M, 50mM Tris-HCl 缓冲溶液(pH 8)
活性: 25unit/min/mg; 单位定义：r Lys-c白蛋酶每分钟酶切1.0 umole T-G-P Lys Nitroanilide的量，反应条件：25℃，波长405nm
Part No. Name Size /pkg
HLS LYS001C rLys-C, Mass Spec Grade 20 μg
HLS HAc001 50 mM Acetic acid buffer 0.5 mL
Description: rLys-C, Mass Spec Grade, specifically hydrolyzes peptide bonds at the carboxyl side of lysine residues. 8 M urea solution is recommended for denaturation. rLys-C can efficiently cleave difficult-to-digest proteins because rLys-C remains its activity under denaturation conditions.
Physical Appearance: Lyophilized powder
Molecular Weight: 27 kDa
Resuspension Buffer (HLS HAc001C): rLys C Resuspension Buffer is composed of 50 mM Acetic acid buffer.
Storage Conditions: Store the lyophilized powder at –20 °C; store reconstituted enzyme at –80 °C.
Shelf life: 24 months at -20 ℃
Stability: rLys-C is maximally active in the pH 8 and reversibly inactivated at pH 3.
Standard In-Solution Protein Digestion:
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For maximum activity, resuspend Mass Spectrum Grade rLys-C in resuspension buffer provided.
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Recommend using 6 - 8 M urea and 50 mM Tris-HCl (pH 8) to denature protein or protein mixture prior to digestion.
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Add rLys-C to the mixture to reach a protein : protease ratio of 25 : 1 (w/w). Mix well and incubate at 37 °C for 3 - 4 hours.
Activity: 25 units/min/mg
rGlu C蛋白酶, 质谱级
rGlu C Protease, MS Grade
订货号： HLS GLU001C
说明: 质谱级rGlu-C蛋白酶特异性水解在谷氨酸或天冬氨酸片段的C端的肽键。该酶适合在溶液中进行消化反应，不推荐用于胶内消化
在碳酸氢铵和醋酸铵缓冲液中，对谷氨酸残基的切割特异性更高；在磷酸盐缓冲液中，谷氨酸和天冬氨酸残基均可被切割
物理形态: 质谱级rGlu-C蛋白酶是冻干粉
分子量: 24kDa.
溶解液：rGlu-C蛋白酶用超纯水溶解
储存条件：冻干粉末存储在-20℃冰箱，再溶解的酶储存在-20℃。有效期见产品标签
保质期：12个月，在-20℃条件下
pH 范围: 在PH 4.0–9.0 时， Glu-C 具有最佳活性
适用范围：rGlu-c蛋白酶特异性水解变性后蛋白质D和E位。
溶液酶解SOP:
50ug rGlu-C 蛋白酶按照 25:1 蛋白和蛋白酶的量(w/w)使用, 在pH8.5下，37°C恒温过夜13-16小时.
活性: 800unit/min/mg；单位定义：rGlu-c白蛋酶每分钟酶切1.0 umole BOC-L-Glutamic Acid a phenyl Ester的量，反应条件：37℃，波长270nm
Part No. Name Size/pkg
HLS GLU001C rGlu-C, Mass Spec Grade 50μg
Description: Endoproteinase rGlu-C, Mass Spec Grade, is a serine proteinase that specifically cleaves peptide bonds at C-terminal to glutamic acid residues. Enzyme specificity to glutamic acid is high in ammonium bicarbonate and ammonium acetate. In phosphate buffers, cleavage occurs at both aspartic and glutamic residues. rGlu-C activity is optimal in the pH range of 4.0 – 9.0. This mass spec grade enzyme can be used alone or in combination with other proteases to generate protein digests.
Physical form: Lyophilized in the presence of trehalose (1.65mg/vial)
Molecular Weight: 24 kDa
Resuspension Buffer: Resuspend rGlu-C powder in double-distilled water from user.
Storage Conditions: Store the lyophilized powder at –4 °C or 20 °C; store reconstituted enzyme solution at -20 °C for up to 8 weeks.
Shelf life: 36 months at -20 ℃
Stability: Maximally active in the pH range of 4-9
In-Solution Protein Digestion Protocol (recommended):
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For maximum activity, resuspend Mass Spectrum Grade rGlu-C with the double-distilled from user.
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Dissolve protein samples with Tris-HCl (pH 8) buffer to a final protein concentration of 50 mM prior to digestion.
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Add rGlu-C proteinase to a protein : proteinase ratio of 20 : 1(w/w). Mix well and incubate at 37 °C for 4 h.
Activity: 800 U/min/mg.
α-Chymotrypsin蛋白酶, 质谱级
α-Chymotrypsin Protease, MS Grade
订货号： HLS CHY001C
说明: 胰凝乳蛋白酶是一种丝氨酸内蛋白酶，可特异性酶切Tyr，Phe，Trp和Leu的C- 末端的肽键。 也有少量的Met，Ala，Asp和Glu的C-末端的肽键裂解。其中的残留胰蛋白酶用TLCK灭活后纯化处理。该蛋白酶用Ca2+离子激活并稳定。
物理形态: 质谱级α -Chymotrypsin蛋白酶是冻干粉
分子量: 25kDa.
溶解液：1mM盐酸(含2uM CaCl2) 100ul复溶
储存条件：冻干粉存储在-20℃冰箱；溶解的酶储存在-20℃有效期1个周。
保质期：冻干粉在-20℃稳定期12个月
pH 范围: 在 PH=7.8时， Chymotrypsin蛋白酶具有最佳活性
适用范围：Chymotrypsin蛋白酶特异性水解变性后蛋白质Y, F, W和L位
溶液酶解SOP:
胰凝乳蛋白酶按照 1:50 蛋白酶和蛋白的量(w/w)使用, 在30°C下于100 mM含10 mM CaCl2，pH 7.8的Tris HCl中进行酶切消化2-12小时。
活性:
70u/mg; 单位定义：在pH 7.8和25°C下,每分钟胰凝乳蛋白酶水解1.0μmol BTEE蛋白的量
Maldi TOF质谱检测: 纯化后的蛋白酶使用基质激光解析质谱分析未见杂质蛋白质峰
Part No. Name Size
HLS CHY001C α-Chymotrypsin 50 μg
Description: Chymotrypsin is a serine endoprotease that specifically cleaves the C-terminal peptide bonds of Tyr, Phe, Trp, and Leu. It also cleaves some C-terminal peptide bonds of Met, Ala, Asp, and Glu. The residual trypsin can be inactivated by TLCK and purified. The protease can be activated and stabilized by Ca2+ ions.
Physical Appearance: Lyophilized powder.
Molecular Weight: 25 kDa.
Resuspension Buffer: Reconstitute lyophilized powder in 100 μL 1 mM hydrochloric acid containing 2 μM CaCl2 (recommended).
Storage Conditions: Store freeze-dried powder at -20 °C refrigerator; store dissolved enzyme at -20 °C, valid for 1 week.
Shelf Life: 24 months at -80 ℃.
Stability: Maximally active in the pH range of 7.8.
In-Solution Protein Digestion Protocol: Use α-chymotrypsin at a protease to protein ratio of 1: 50 (w/w), and perform digestion in 100 mM Tris-HCl (pH 7.8) containing 10 mM CaCl2 at 30 °C for 2 - 12 hours.
Tips:
1. The protease will self-hydrolyze if the water bath temperature is higher than 37 °C.
2. Preferably < 1M urea or guanidine hydrochloride concentration during the sample digestion.
Activity: 70 U/mg.
rTrypsin 3.0蛋白酶, 质谱级
rTrypsin 3.0 Protease, MS Grade
订货号： HLS rTRY001C
说明: rTrypsin 3.0胰蛋白酶是重组表达的金属蛋白酶。超快速特异性裂解精氨酸(R)和赖氨酸(K)的C端肽键。该蛋白酶经过基因序列重新设计并甲基化修饰处理，具有超快速酶解蛋白，不自降解和高活性等特点。用于蛋白表征，单细胞组学和大队列蛋白组学研究
物理形态: 冻干粉含20ug海藻糖
分子量: 23.7kDa.
溶解液: 20ul 1mmol 盐酸溶解即浓度1ug/ul
存储条件: –20 °C冰箱冷冻.
保质期：24 个月 在 –20°C冰箱
pH值范围: rTrypsin 3.0蛋白酶在pH7-9活性最佳
酶切方法:
-适用酶切缓冲溶液：20mmol Tris，50mmolABC 或 HEPES 缓冲液;， pH值在7-9范围内
-变性蛋白样品，按照1：50蛋白酶和蛋白量使用，酶切反应蛋白浓度控制在0.5ug/ul, 45°C干浴孵育30min
活性: 15000unit/mg; 单位定义：在pH 7.8和25°C下, 波长253nm，每分钟胰蛋白酶水解1.0μmol BAEE蛋白的量
Part No. Name Size/pkg
HLS rTRY001C rTrypsin 3.0, Mass Spec Grade 20 μg
Description: rTrypsin 3.0 is a recombinant protease that specifically hydrolyzes peptide bonds at the carboxyl side of lysine and arginine residues. Engineered with a redesigned gene sequence and methylated lysines, it offers ultra-fast protein hydrolysis, high activity, and no self-degradation. rTrypsin 3.0 is ideal for protein characterization, single-cell proteomics, and large cohort proteomics studies.
Physical Appearance: Lyophilized powder with 20 μg Trehalose
Molecular Weight: 23.7 kDa
Resuspension Buffer (HLS HAC001C): Dissolved in 40 μL of 50 mmol acetic acid, concentration 0.5 μg/uL
Storage Conditions: Store the lyophilized powder at –20 °C; store reconstituted enzyme at –80 °C.
Shelf life: 24 months at -20 ℃
Stability: rTrypsin 3.0 is maximally active in the pH range of 7–9
Usage Notes:
1. Suitable enzyme digestion buffers: 20 mmol Tris, 50 mmol ABC, or HEPES buffer; pH 7-9.
2. Use a 1:50 protease-to-protein ratio with a protein concentration of 0.5 µg/µL. Incubate at 37°C in a dry bath for 30 minutes.
Activity: rTrypsin: 15,000 units/mg. Unit Definition: At pH 7.8 and 25°C, 1.0 μmol of BAEE protein is hydrolyzed per minute at 253 nm.
rTrypsin/rLys C复合蛋白酶, 质谱级
rTrypsin/rLys C Mixer Protease, MS Grade
订货号： HLS rTRYLYSC
说明: rTrypsin 3.0胰蛋白酶和rLys C蛋白酶是重组表达的金属蛋白酶。rTrypsin 蛋白酶超快速特异性裂解精氨酸(R)和赖氨酸(K)的C端肽键; rLys C蛋白酶快速特异性裂解赖氨酸(K)的C端肽键。由于胰蛋白酶不能以相同的速率酶解精氨酸和赖氨酸，造成许多赖氨酸的漏切; 同时有的赖氨酸有PTM修饰或C端紧邻强疏水性氨基酸(例如P), 也会造成漏切问题。rTrypsin/rLys C复合蛋白酶适合处理难酶解消化的复杂蛋白样品。用于蛋白表征，单细胞组学和大队列蛋白组学研究。
物理形态: 冻干粉含67ug海藻糖
分子量: rTrypsin 23.7kDa; rLys C 27kDa
溶解液: 40ul 50mmol 醋酸溶解即浓度0.5ug/ul
存储条件: –20 °C冰箱冷冻.
保质期：24 个月 在 –20°C冰箱
pH值范围: 复合蛋白酶在pH7-9活性最佳
酶切方法:
-适用酶切缓冲溶液：20mmol Tris，50mmolABC 或 HEPES 缓冲液;， pH值在7-9范围内
-变性蛋白样品，按照1：50蛋白酶和蛋白量使用，酶切反应蛋白浓度控制在0.5ug/ul, 37°C干浴孵育30min
活性: rTrypsin: 15000unit/mg; 在pH 7.8, 25°C条件下, 波长253nm，每分钟胰蛋白酶水解1.0μmol BAEE蛋白的量
rLys C: 25unit/mg; pH7.8，25°C条件下, 波长405nm，每分钟rLys C水解1.0μmol T-G-P lys Nitroanilid的量
Part No. Name Size/pkg
HLS rTRYLYSC rTrypsin/rLys C, Mass Spec Grade 20 μg
Description: The rTrypsin 3.0 and rLys C enzyme mix combines two recombinant proteases for efficient peptide bond hydrolysis. rTrypsin 3.0 cleaves at the carboxyl side of lysine and arginine residues, while rLys C targets lysine residues specifically. This combination overcomes issues with missed cleavages due to rTrypsin’s slower digestion of lysine and arginine, PTM modifications on lysines, or hydrophobic C-termini (e.g., proline). The rTrypsin and rLys C mix is ideal for protein characterization, single-cell proteomics, and large cohort proteomics studies.
Physical Appearance: Lyophilized powder with 67 µg Trehalose
Molecular Weight: rTrypsin 23.7kDa; rLys C 27kDa
Resuspension Buffer (HLS HAC001C): Dissolve in 40 µL of 50 mmol acetic acid, yielding a concentration of 0.5 µg/µL.
Storage Conditions: Lyophilized powder: –20°C; Reconstituted enzyme: –80°C
Shelf life: 24 months at -20 ℃
Stability: rTrypsin 3.0/rLys C is maximally active in the pH range of 7–9
Usage Notes:
1. Suitable digestion buffers: 20 mmol Tris, 50 mmol ABC, or HEPES buffer (pH 7–9).
2. Use denatured protein samples at a 1:50 protease-to-protein ratio. Maintain a protein concentration of 0.5 µg/µL and incubate at 37°C for 30 minutes
Activity:
• rTrypsin: 15,000 units/mg. Unit Definition: 1.0 µmol BAEE hydrolyzed per minute at pH 7.8, 25°C, 253 nm.
• rLys C: 25 units/mg. Unit Definition: 1.0 µmol T-G-P Lys Nitroanilid hydrolyzed per minute at pH 7.8, 25°C, 405 nm.
Arg C蛋白酶, 质谱级
Arg C Protease, MS Grade
订货号： HLS ARG001C
说明: Arg C蛋白酶是一种内肽酶，可在精氨酸残基的 C 端切割，含脯氨酸旁边的位点。可以与其他蛋白酶结合使用，用于蛋白组学质谱分析。
物理形态: 质谱级Arg C蛋白酶是冻干粉(含0.25mg海藻糖, 0.1µmol半胱氨酸和2umol CaCl 2）
分子量: 45kDa.
溶解液：1mM盐酸(含2mM CaCl2) 10复溶
储存条件：冻干粉存储在-20℃冰箱；溶解的酶储存在-20℃有效期1个月。
保质期：冻干粉在-20℃稳定期12个月
pH 范围: 在 pH 7.6–7.9 时活性最佳。
适用范围：Arg C蛋白酶特异性酶切变性后蛋白质R c-bond肽键