蛋白酶-C 产品/ Endoproteinase-C Products

 

Trypsin 蛋白酶, 质谱级

 

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

 

Trypsin, Mass Spec Grade

 

Part No.                    Name                                                           Size/pkg

HLS TRY001C         Trypsin, Mass Spec Grade1mM            100µg

HLS HCL001           hydrochloric acid buffer                         0.5ml

 

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, auto- proteolysis can result in the generation of pseudo trypsin, which has been shown to exhibit chymotrypsin-like specificity.Hualishi Scientific 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-hydrolysis and retain higher the activity of general trypsin.

Physical form: mass spectrometry modified trypsin is supplied by lyophilized powder.

Molecular Weight: 23kDa.

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 –70°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 Mass Spectrum Grade Modified Trypsin in the Trypsin Resuspension Buffer provided, and heat at 30°C for 15 minutes before use.

-Thaw the reconstituted trypsin at room temperature, placing on ice immediately after thawing. Remove the amount of trypsin needed, then refreeze the unused portion at -70℃. To maintain maximum product activity, limit the number of freeze-thaw cycles to five or dispense into single-use aliquots after resuspending.

-A protease:protein ratio of 1:100 to 1:25(w/w) is recommended for protein identification or PTM for 13-16 hours at 37°C

rLys-C蛋白酶, 质谱级
 
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

 

rLys-C, Mass Spec Grade

 

Part No.                  Name                                         Size/pkg

HLS LYS001C        rLys-C, Mass Spec Grade         20µg

HLS HAc001          50mM Acetic acid buffer            0.5ml

 

Description: rLys-C, Mass Spec Grade, specifically hydrolyzes peptide bonds at the carboxyl side of lysine residues. Recommended for use in solutions with up to 8M Urea. rLys-C can efficiently cleave difficult-to-digest proteins due to the ability of Lys-C to remain active under denaturing conditions.

Physical form: supplied by lyophilized powder.

Molecular Weight: 27kDa.

Resuspension Buffer(HLS HAc001C): rLys-C Resuspension Buffer is composed of 50mM Acetic acid( PH=3.0).

Storage Conditions: Store the lyophilized powder at –20°C. Store reconstituted enzyme at –70°C for up to 30 days. To retain maximum activity, do not freeze and thaw more than once.

Shelf life:24 months at -20℃

Stability: rLys-C is maximally active in the pH range of 8 and reversibly inactivated at pH 3.

Activity:  25unit/min/mg;

Unit Definition: One unit is the amount of Mass spectrum Grade rLys-C will hydrolyze 1.0 umol of N-p-Tosyl-Gly-Pro-Lysp-Nitroanilide per minute at PH=7.7 at 25°C, A405, Light path=1cm

rGlu-C蛋白酶, 质谱级

 

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

 

rGlu-C, Mass Spec Grade

 

Part No.                      Name                                            Size/pkg  

HLS GLU001C           rGlu-C, Mass Spec Grade            50µg

HLS WAT001             double-distilled water                     n/a

 

Description: Endoproteinase rGlu-C, Mass Spec Grade, is a serine proteinase which specifically cleves peptide bonds C-terminal to glutamic acid residues. In ammonium bicarbonate and ammonium acetate, the enzyme specificity is higher at the glutamic residues. In phosphate buffers, cleavage occurs at the aspartic and glutamic residues . Glu-C activity is optimal in the pH range of 4.0–9.0. This sequencing grade enzyme can be used alone or in combination with other proteases to produce protein digests.

 

Physical form: Lyophilized in the presence of trehalose (1.65mg/vial)

Molecular Weight: 24kDa.

Resuspension Buffer(HLS WAGT001): rGlu-C Resuspension Buffer is double-distilled from user.

Storage Conditions: Store the lyophilized powder at –20°C or 4°C, Reconstituted enzyme solution can be stored at –20°C for 8 weeks.

Shelf life:36 months at -20℃

Stability: rGlu-C is maximally active in the pH range of 4-9 .

In-Solution Protein Digestion:

-For maximum activity, resuspend Mass Spectrum Grade rGLu-c in the double-distilled from user.

 

-we recommend using Tris-HCl (pH 8) buffer to a protein to a final concentration of 50mM, before the digestion.

-Add rGlu-C to protein at a 20:1 protein:protease ratio (w/w), mix and incubate for 16 hours at 37°C

Activity:  800unit/min/mg;

Unit Definition: One unit is the amount of Mass spectrum Grade rGlu-C will hydrolyze 1.0 umol of N-t-Boc--L-glutamic acid a-phenyl easter

per minute at PH=7.8 at 37°C, A270, Light path=1cm

​© 2020年02月02日 北京华利世科技|北京生夏蛋白技术  版权所有.  

E-mail: yu.feng@.coowins.com | Phone: +86 139 1067 9882

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