上海金畔生物代理MiTeGen品牌蛋白结晶试剂耗材工具等，我们将竭诚为您服务，欢迎访问MiTeGen官网或者咨询我们获取更多相关MiTeGen品牌产品信息。
Shelved Storage cane for MiTeGen 2nd Generation Cryo-EM Pucks. Holds up to 10 pucks. Compatible with HC34 and VHC35.
上海金畔生物代理MiTeGen品牌蛋白结晶试剂耗材工具等，我们将竭诚为您服务，欢迎访问MiTeGen官网或者咨询我们获取更多相关MiTeGen品牌产品信息。
Shelved Storage cane for MiTeGen 2nd Generation Cryo-EM Pucks. Holds up to 10 pucks. Compatible with HC35.
New England Biolabs supplies a 10X reaction buffer with all of its enzymes. At a 1X concentration this reaction buffer assures optimal activity of the enzyme.
Storage Temperature
-20°C
20 mM Tris-HCl
15 mM MgCl2
50 mM NaCl
0.1 mg/ml BSA
0.15% Triton® X-100
pH 7.5@25°C
产品于 2020 年 1 月 2 日停产，产品组分可单独购买
SNAP-Cell 启动试剂盒（停产，组分可单独购买）
SNAP-Surface 启动试剂盒
在免疫沉淀实验流程中富集经 m6A 修饰的 RNA
EpiMark N6-甲基化腺嘌呤富集试剂盒包含一个 N6– 甲基腺苷（m6A）兔源单克隆抗体。试剂盒中含有 2 个对照RNA，一个含有 m6A 修饰（Gaussia 荧光素酶），另一个不含 m6A 修饰（Cypridina 荧光素酶）用于监测富集和消耗程度。其中，GLuc RNA 对照在转录过程中需 20% 的 m6ATP 和 80% 的 ATP。
该试剂盒可用于在免疫沉淀实验流程中富集经 m6A 修饰的 RNA，可用于 NGS 或 RT-qPCR 等下游实验。在片段化的 RNA 样本中，经修饰的 RNA 可与 N6– 甲基腺苷抗体结合，从而被与抗体结合的 Protein G 磁珠富集。再经多轮洗涤与纯化步骤后，富集的 RNA 洗脱于无核酸酶水中，可用于进一步的分析。
– N6–甲基腺苷抗体
1 set of this bundle includes 2,000,000 units of O-Glycosidase and 2,000 units of Neuraminidase.
O-Glycosidase, also known as Endo-α-N-Acetylgalactosaminidase, catalyzes the removal of
Core 1 and Core 3 O-linked disaccharides from glycoproteins.
Neuraminidase is the common name for Acetyl-neuraminyl hydrolase ( Sialidase ). This Neura-minidase catalyzes the hydrolysis of α2-3, α2-6, α2-8 linked N-acetyl-neuraminic acid residues
from glycoproteins and oligosaccharides.
O-Glycosidase is cloned from Enterococcus faecalis and expressed in E.coli (1). Neuraminidase
is cloned from Clostridium perfringens (2) and overexpressed in E. coli (3).
The following reagents are supplied with this product:
Unit Definition
One unit of O-Glycosidase is defined as the amount of enzyme required to remove 0.68 nmol of O-linked disaccharide from 5 mg of Neuraminidase digested, non-denatured fetuin in 1 hour at 37°C in a total reaction volume of 100 µl (1 unit of both O-Glycosidase and PNGase F will remove equivalent molar amounts of O-linked disaccharides and N-linked oligosaccharides, respectively).
Non-denaturing Unit Definition of O-Glycosidase:
Two fold serial dilutions of O-Glycosidase are added to a reaction mixture of 5 mg of Neura-minidase digested fetuin with 1 X GlycoBuffer 2. The reaction is then incubated at 37°C for 1 hour. O-linked disaccharide carbohydrates are determined by Morgan and Elson Assay (4). Note: Under denaturing conditions the enzyme activity is increased two-fold. This observation is substrate dependent.
Unit Definition of Neuraminidase:
One unit of Neuraminidase is defined as the amount of enzyme required to cleave > 95% of the terminal α-Neu5Ac from 1 nmol Neu5Acα2-3Galβ1-3GlcNAcβ1-3Galβ1-4Glc-7-amino-4-methyl- coumarin (AMC), in 5 minutes at 37°C in a total reaction volume of 10 µl.
1X Glycoprotein Denaturing Buffer
0.5% SDS
40 mM DTT
1X NP-40
1% NP-40 in MilliQ-H2O
1X GlycoBuffer 2
Incubate at 37°C
1X GlycoBuffer 2:
50 mM sodium phosphate
pH 7.5 @ 25°C
Companion Products
α2-3,6,8 Neuraminidase
O-Glycosidase
PNGase F
PNGase F (Glycerol-free)
Endo H
Endo Hf
1.Since O-Glycosidase is inhibited by SDS, it is essential to have NP-40 in the reaction mixture. It is not known why this non-ionic detergent counteracts the SDS inhibition at the present time. Double digest with Endo H must have NP-40 present (NP-40 does not inhibit Endo H).
2.To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
1.Koutsioulis, D., Landry, D. and Guthrie, E.P. (2008). Glycobiology. 18, 799-805.
2.Roggentin, P. et al. (1988). FEBS Lett. 238, 31-34.
3.Guan, C. unpublished observations. New England Biolabs.
4.Morgan, W.T.J. and Elson, L.A. (1934). Biochem. J. 28, 988-995.
NEB代理 , Buffers , T4 Polynucleotide Kinase Reaction Buffer,#B0201S
NEB代理 , Buffers , T4 Polynucleotide Kinase Reaction Buffer,#B0201S
去除糖蛋白中的 N-连接高甘露糖
无糖苷外切酶污染，无蛋白水解活性。
酶活性不受 SDS 影响。
要使天然糖蛋白去糖基化，可能需要增加酶量并延长温育时间。
