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重组白蛋白, 分子生物学级 |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

重组白蛋白（rAlbumin）可以作为 BSA 的替代品，能够防止酶附着在反应试管和移液管管壁上，还能在温育过程中稳定某些蛋白质。欢迎咨询定制配方重组白蛋白。

产品类别:
Restriction Endonuclease Buffers & Diluents Products,
Buffers Products

应用:
Restriction Enzyme Digestion,
Restriction Enzyme Digestion

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • B9200S     -20    
        Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml

  • 特性和用法

    贮存溶液

    20 mM Tris-HCl
    100 mM KCl
    0.1 mM EDTA
    50% Glycerol
    pH 8 @ 25°C

  • 相关产品

    相关产品

    • NEBuffer 4
    • Nuclease-free Water

FAQs & 问题解决指南

  • FAQs

    1. What is the source of Recombinant Albumin, Molecular Biology Grade?
    2. What is the buffer composition of Recombinant Albumin, Molecular Biology Grade?
    3. Why would I want to add Recombinant Albumin, Molecular Biology Grade to my reaction?
    4. Can I use Recombinant Albumin, Molecular Biology Grade as a molecular weight marker?
    5. Can I use Recombinant Albumin, Molecular Biology Grade as a concentration standard?
    6. What is the difference between BSA Molecular Grade and Recombinant Albumin, Molecular Biology Grade?
    7. Is Recombinant Albumin, Molecular Biology Grade acetylated?

O-糖蛋白酶（IMPa） |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

O-糖蛋白酶（IMPa）又称免疫调节蛋白酶（IMPa），具有广泛特异性，是一种克隆自铜绿假单胞菌（Pseudomonas aeruginosa）的 O-糖蛋白酶。该酶可以识别糖蛋白或糖肽链中的粘蛋白型 O-糖链（包括分支结构和唾液酸化结构），并将肽键切割为糖基化丝氨酸或苏氨酸残基（在 N 末端切割）。糖蛋白质组学实验流程可使用该酶来绘制 O-糖基化残基图谱、分析各糖位点上 O-糖链的结构特征。O-糖蛋白酶（IMPa）具有 6xHis 标签，可使用镍亲和树脂轻松从反应中将其去除。

 

特异性：

O-糖蛋白酶（IMPa） |

产品来源

该酶克隆自铜绿假单胞菌（Pseudomonas aeruginosa），在 E. coli 中表达。

产品类别:
Protein Tools Products,
Proteases Products,
Glycobiology Products,

Proteome Analysis Products

应用:
Glycomics and glycoproteomics,
Protein Digestion,
Glycobiology & Proteomics,

Proteomics,

Glycoprotein Analysis

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P0761S     -20    
        O-Glycoprotease (IMPa) P0761SVIAL -20 1 x 200 reactions 1,000 units/ml

  • 特性和用法

    单位定义

    1 单位 O-糖蛋白酶（IMPa）指在 20 µl 总反应体系中（20 mM Tris-HCl，pH 8.0），37℃ 条件下，2 小时能切割 90% 以上 2 µM FAM 标记的 O-糖肽链所需的酶量。

    贮存溶液

    20 mM Tris-HCl
    100 mM NaCl
    pH 7.5 @ 25°C

    热失活

    95°C for 10 minutes

    分子量

    理论上的: 97 kDa

    单位活性检测条件

    将两倍稀释的 O-糖蛋白酶（IMPa）、2 μM FAM 标记的 O-糖肽链和 20 mM Tris-HCl（pH 8.0）在 20 µl 反应体系中温育。将反应混合液 37℃ 温育 2 小时。通过 CE 分析确定反应产物的分离情况。

  • 优势和特性

    Features

    • O-糖基化位点图谱构建
    • O-糖链结构确定
    • 能够表明含 O-连接糖链的简单和复杂糖蛋白的特征
    • 在含或不含唾液酸的情况下，有效切割糖蛋白，无需神经氨酸苷酶处理
    • 200 次反应（一管）足以切割高达 2 mg 的糖蛋白

  • 相关产品

    相关产品

    • Trypsin-ultra™, Mass Spectrometry Grade
    • p0733-o-glycosidase
    • O-糖苷酶 & 神经氨酸苷酶套装
    • p0709-pngase-f-glycerol-free-recombinant

  • 注意事项

    1. O-糖蛋白酶（IMPa）可消化多种含有粘蛋白型 O-糖链（含或不含唾液酸）的糖蛋白和糖肽链，包括 Tn 抗原（GalNAc-α-Ser/Thr）。
    2. 糖蛋白可以先用 O-糖蛋白酶（IMPa）消化，再用胰蛋白酶-ultra（质谱级）（NEB #P8101）消化。
    3. O-糖蛋白酶（IMPa）活性的最适反应缓冲液是 20 mM Tris-HCl（pH 8.0）。O-糖蛋白酶（IMPa）的最适 pH 值范围是 7.0-8.0。pH 值低于 7.0 或高于 8.0 时，酶活性显著降低。

操作说明、说明书 & 用法

  • 操作说明

    1. O-Glycoprotease (IMPa) On-filter Cleavage Denaturing Reaction Protocol (NEB #P0761)
    2. O-Glycoprotease (IMPa) Denaturing Reaction Protocol (NEB #P0761)
    3. O-Glycoprotease (IMPa) Non-Denaturing Reaction Protocol (NEB #P0761)

  • 应用实例

    • Comprehensive structural and positional profiling of intact complex O-glycans in biologic drugs using O-Glycoprotease (IMPa)

FAQs & 问题解决指南

  • FAQs

    1. Can an O-Glycoprotease (IMPa) reaction be scaled linearly to accommodate digestion of greater than 10 μg of glycoprotein?
    2. Is O-Glycoprotease (IMPa) active in the presence of reducing agents or detergents?
    3. Is O-Glycoprotease (IMPa) active in buffers other than 20 mM Tris-HCl pH 8.0?
    4. Is O-Glycoprotease (IMPa) able to efficiently cleave clustered O-glycosylated sites?
    5. I see minimal cleavage of my glycoprotein after treatment with O-Glycoprotease (IMPa). How can I optimize the reaction?
    6. Is it necessary to first remove sialic acids from the substrate prior to digest with O-Glycoprotease (IMPa)?
    7. Is O-Glycoprotease (IMPa) active on TF-antigen and Tn-antigen?
    8. Does O-Glycoprotease (IMPa) require metal ions for activity?

Endo H |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

Endo H |
 
Endoglycosidase H is a recombinant glycosidase which cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N-linked glycoproteins.

Endo H | Endo H |

60 µg of RNase B was incubated with 3,000 units of Endo H or Endo Hf under standard assay conditions

Endo H |
Aliquots were removed at various time points and measured for released carbohydrate. [Dubois et al. (1956) Anal. Chem. 28, 350-356].
Mobility Shift Analysis
Endo H |
1 unit of Endo H, Endo Hf or PNGase F was incubated per 10 µg of RNase B under standard assay conditions. At various time points, aliquots were removed and analyzed on a 10-20% SDS-PAGE gel for carbohydrate (CHO) release. 1 unit is defined as the amount of enzyme required to remove >95% of the carbohydrate from 10 µg of RNase B in one hour at 37°C.

产品来源

Cloned from Streptomyces picatus (2) and overexpressed in E.coli (3).

产品类别:
Endoglycosidases Products,
Proteome Analysis Products

应用:
Expression Systems,
Glycan Sequencing,
Proteomics,

Recombinant Glycoprotein Expression,

Glycoprotein Analysis

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P0702S     -20    
        Endo H P0702SVIAL -20 1 x 0.02 ml 500,000 units/ml
        GlycoBuffer 3 B1720SVIAL -20 1 x 1 ml 10 X
        Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X
    • P0702L     -20    
        Endo H P0702LVIAL -20 1 x 0.1 ml 500,000 units/ml
        GlycoBuffer 3 B1720SVIAL -20 1 x 1 ml 10 X
        Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10 µl (10 NEB units = 1 IUB milliunit). 

    Unit Definition Assay: 
    10 µg of RNase B are denatured with 1X Glycoprotein Denaturing Buffer at 100°C for 10 minutes. After the addition of 1X GlycoBuffer 3, two-fold dilutions of Endo H are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products is visualized by SDS-PAGE.

    1X Glycoprotein Denaturing Buffer
    0.5% SDS
    40 mM DTT

    反应条件

    1X GlycoBuffer 3
    Incubate at 37°C

    1X GlycoBuffer 3
    50 mM sodium acetate
    (pH 6 @ 25°C)

    贮存溶液

    20 mM Tris-HCl
    50 mM NaCl
    5 mM EDTA
    pH 7.5 @ 25°C

    热失活

    75°C for 10 minutes

    分子量

    实际: 29 kDa

  • 优势和特性

    应用特性

    • Removal of high mannose N-glycans from glycoproteins

  • 相关产品

    相关产品

    • p0703-endo-hf
    • p0704-pngase-f
    • p0705-pngase-f-glycerol-free
    • RNase B（对照底物）
    • p0706-remove-it-pngase-f
    • Endo S
    • 糖苷内切酶 D

  • 注意事项

    1. Enzymatic activity is not affected by SDS.
    2. To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
    3. Activity at different temperatures (measured after a 1 hour incubation of glycosidase and denatured RNase B at the given temperature): 37°C – 100%; 30°C – 65%; 25°C – 40%; 17°C – 25% and  2°C – 0%.
    4. Typical reaction conditions: Please see FAQs.

  • 参考文献

    1. Maley, F. et al. (1989). Anal. Biochem. 180, 195-204.
    2. Robbins, P. et al. (1984). J. Biol. Chem. 259, 7577-7583.
    3. Guan, C. and Wong,S. New England Biolabs, unpublished observations.

操作说明、说明书 & 用法

  • 操作说明

    1. Endo H/Endo Hf Protocol
    2. Protocol for Endo H/Hf Non-Denaturing P0702 and P0703

工具 & 资源

  • 选择指南

    • Endoglycosidase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. What is the difference between PNGase F, Endo H and O-Glycosidase?
    2. What is the difference between Endo H and Endo Hf?
    3. I tried the Endo H/Hf on my glycoprotein and it failed. What could be the problem?
    4. How much Endo H/Endo Hf should I use?
    5. Is EndoH/ Endo Hf inhibited by SDS?
    6. What are the typical reaction conditions for Endo H?
    7. Are Protease Inhibitors acceptable for use in an Endo H/Hf reaction?
    8. What are Glycosidases and their uses?
    9. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
    10. What is a good endoglycosidase substrate?
    11. Do detergents inhibit exoglycosidases/endoglycosidases?

  • 实验技巧

    You can use this enzyme under native or denaturing conditions

    Under native conditions we recommend adding more enzyme and using longer incubation times

    Enzymatic activity is not affected by SDS

    A good positive control substrate is RNase B

Endo S |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

Endo S |
 
Endo S is an endoglycosidase with a uniquely high specificity for removing N-linked glycans from the chitobiose core of the heavy chain of native IgG (1). Endo S is tagged with a chitin binding domain (CBD) for easy removal from a reaction and is supplied glycerol free for optimal performance in HPLC and MS intensive methods.

产品来源

Endo S is cloned from Streptococcus pyogenes and overexpressed as a fusion to the chitin binding domain in E. coli.

特异性

Detailed Specificity
X = protein, peptide, Asparagine, or free glycan, as Endo S does not have a strict peptide requirement for activity. Endo S is active on a substrate with or without core α(1-6)fucosylation as well as a with or without a bisecting N-acetylglucosamine. Triantennary and tetrantennary sialyated or asialo glycans are not a substrate for Endo S (1).

产品类别:
Endoglycosidases Products,
Proteome Analysis Products

应用:
Expression Systems,
Glycan Sequencing,
Proteomics,

Recombinant Glycoprotein Expression,

Glycoprotein Analysis

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P0741S     4    
        Endo S P0741SVIAL 4 1 x 0.03 ml 200,000 units/ml
        GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
    • P0741L     4    
        Endo S P0741LVIAL 4 1 x 0.15 ml 200,000 units/ml
        GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 5 μg of native mouse monoclonal IgG in 1 hour at 37°C in a total reaction volume of 10 µl.

    反应条件

    1X GlycoBuffer 1
    Incubate at 37°C

    1X GlycoBuffer 1
    5 mM CaCl2
    50 mM sodium acetate
    (pH 5.5 @ 25°C)

    贮存溶液

    20 mM Tris-HCl
    50 mM NaCl
    5 mM EDTA
    pH 7.5 @ 25°C

    热失活

    55°C for 10 minutes

    分子量

    实际: 136000 daltons

    单位活性检测条件

    5 µg of IgG in 1X GlycoBuffer 1 are incubated with two-fold dilutions of Endo S for 1 hour at 37°C. Separation of reaction products is visualized by SDS-PAGE.

  • 相关产品

    相关产品

    • 几丁质磁珠
    • 6 孔磁性分离架
    • 12 孔磁性分离架
    • p0706-remove-it-pngase-f
    • 糖苷内切酶 D
    • IdeZ Protease (IgG-specific)
    • p6043-rapid-pngase-f-antibody-standard
    • p0711-rapid-pngase-f-non-reducing-format

  • 注意事项

    1. Recommended storage temperature is 4°C,avoid repeat freeze-thaw cycles.
    2. Removal of Endo S from thedeglycosylation reaction can be scaled uplinearly with larger volumes of chitin magneticbeads.
    3. The binding capacity of Chitin Magnetic Beads is 1 mg of CBD-tagged protein / mL of beads.

  • 参考文献

    1. Bielik, A., McLeod, E. and Magnelli, P., New England Biolabs,Inc., unpublished results.. Unpublished observation

操作说明、说明书 & 用法

  • 操作说明

    1. Reaction Conditions for Endo S (P0741)
    2. Endo S Removal Magnetic Chitin Bead Protocol (P0741)

  • 应用实例

    • AppNote_Glycan_Analysis_of_Murine_IgG_by_Enzymatic_Digestion_with_Endo_S_and_PNGase_F
    • Glycan Analysis of Murine IgG by Enzymatic Digestion with Endo S and PNGase F Followed by Mass Spectrometric Analysis

工具 & 资源

  • 选择指南

    • Endoglycosidase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. Is Endo S tagged?
    2. What is the difference between PNGase F and Endo S?
    3. How much Endo S should I use to deglycosylate a glycoprotein under native conditions?
    4. What is the preferred substrate for Endo S?
    5. What are the typical reaction conditions for Endo S?
    6. How do I eliminate Endo S from a reaction?
    7. What is the binding capacity of the Magnetic Chitin Beads used to remove Endo S?
    8. Is Endo S compatible with downstream analysis such as HPLC and Mass Spectrometry?
    9. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
    10. What are Glycosidases and their uses?