上海金畔生物科技有限公司代理日本同仁化学 DOJINDO代理商全线产品,欢迎访问官网了解更多信息
C16H17N2NaO9S
436.37
上海金畔生物科技有限公司代理日本同仁化学 DOJINDO代理商全线产品,欢迎访问官网了解更多信息
C16H17N2NaO9S
436.37
上海金畔生物科技有限公司代理日本同仁化学 DOJINDO代理商全线产品,欢迎访问官网了解更多信息
C18H22N3NaO8S3
527.57
关联产品
实验工具|稀释计算器|摩尔浓度计算器
上海金畔生物科技有限公司代理日本同仁化学 DOJINDO代理商全线产品,欢迎访问官网了解更多信息
分子式:
C19H27N4NaO9S4
分子量:
606.69
关联产品
上海金畔生物科技有限公司代理日本同仁化学 DOJINDO代理商全线产品,欢迎访问官网了解更多信息
试剂盒内含
产品概述
水溶性活性酯型生物素化试剂:使用生物素(AC5)2Sulfo-OSu用于生物素标记蛋白质游离氨基的试剂盒。它带有用于标记的LVDS3缓冲粉,用于凝胶过滤的柱和用于柱洗脱的PBS片剂,可以纯化标记的蛋白。由于是单次注射型(10毫克x4瓶),因此可以标记4种不同类型的蛋白质。可以通过改变添加的生物素-(AC5)2 Sulfo-OSu的量来控制标记率。
*标记蛋白的量为1-5 mg。
原理
操作步骤
使用注意事项:
1. 本试剂盒请在0-5 ℃储存。
2. Biotin-(AC5)2Sulfo-OSu溶液需要现配现用 (由于Biotin-(AC5)2Sulfo-OSu易水解,尽量避免在水溶液的状态保存)。
3. 标记好的蛋白质请在0-5 ℃保存 (加入0.1%的叠氮钠等防腐剂)。
使用步骤:
1. 配制溶液
1) NaHCO3缓冲液
在含有NaHCO3粉末的容器中加入10ml超纯水溶解。
2) PBS缓冲液
在100ml容量瓶中加入1片PBS,先用少量超纯水溶解后,再用超纯水补充至100ml,制成10mmol/l的PBS缓冲液。
3) 蛋白质溶液
在样品管中精确称量1.0-5.0mg蛋白质并记录称量值后,用移液器加入500μl操作步骤1)中的NaHCO3缓冲液。盖上盖子后,用漩涡振荡器等搅拌溶解蛋白质。
4) Biotin-(AC5)2Sulfo-OSu溶液
在1支含有Biotin-(AC5)2Sulfo-OSu的管子中加入适量纯水溶解。为了控制标记率,根据不同的标记对象,请参考表1调整Biotin-(AC5)2Sulfo-OSu溶液的浓度及加入量。
*由于Biotin-(AC5)2Sulfo-OSu易水解,溶解后请尽快标记。
2. 生物素标记操作方法
1) 请参考表1在配制好的蛋白质溶液中加入合适浓度和用量的Biotin-(AC5)2Sulfo-OSu溶液。
2) 盖紧盖子充分混合后,在25 ℃水浴恒温摇床中培养2h。
表1 Biotin-(AC5)2Sulfo-OSu标记各种蛋白质的标记率
以上数据为本公司实际检测的结果,如实验条件变动,
结果有可能改变。标记率是根据HABA法测得。
a) 1mol蛋白质:生物素的摩尔数
b) 1mol蛋白质上所结合的生物素的摩尔数
3. 蛋白质标记后的凝胶过滤纯化
1) 打开层析柱上的盖子,弃去填充液。
2) 打开层析柱出口的盖子,用PBS分数次洗脱,每根层析柱收集大约10ml的洗脱液,使凝胶平衡。
3) 用移液器吸取500μl标记好的蛋白质溶液加入到层析柱中,弃去此时的流出液。
4) 在层析柱的出口处放置一个2ml的样品管,在层析柱中加入1ml PBS,收集流出的生物素标记蛋白质。
*经过层析,生物素标记蛋白质的最终浓度为标记时的1/2。
按照下列公式计算生物素标记蛋白质的最终浓度:
A (mol/l) = (X/MWprotein) × (1,000/Y) × 0.5
X:蛋白质的重量
MWprotein:蛋白质的分子量
Y:溶解蛋白质的NaHCO3缓冲液体积
常见问题Q&A
Q1:如何存储标记的蛋白质? |
A:将标记的蛋白质存放在冰箱中。另外,添加0.1%叠氮化钠作为防腐剂。 |
Q2:生物素标记试剂在水溶液中稳定吗? |
A:试剂盒中使用的生物素-(AC5)2 Slufo-OSu处于水溶液中时,由于它会水解,因此无法存储在水溶液中。使用前请做好准备。另外,即使在粉末状态下,也要注意由于吸湿引起的劣化。 |
参考文献
1) J. Wormmeester, F. Stiekema and C. Groot, “Immunoselective Cell Separation”, Methods Enzymol., 1990, 184, 314. |
2) J. J. Leary and D. J. Ward, “Rapid and Sensitive Colorimetric Method for Visualizing Biotin-labeled DNA Probes Hybridized to DNA or RNA Immoilized on Nitrocelulose: Bio-blots”, Proc. Natl. Acad. Sci. USA, 1983, 80, 4045. |
3) W. T. Lee and D. H. Conrad, “The Murine Lymphocyte Receptor for IgE II. Characterization of the Multvalent Nature of the B Lymphocyte Receptor for IgE”, J. Exp. Med., 1984, 159, 1790. |
4) D. R. Gretch, M. Suter and M. F. Stinski, “The Use of Biotinylated Monoclonal Antibodies and Streptavidin Affinity Chromatography to Isolate Herpesvirus Hydorophobic Proteins or Glycoproteins”, Anal. Biochem., 1987, 163, 270. |
5) M. Shimkus, J. Levy and T. Herman, “A Chemically Cleavable Biotinylated Nucleotide: Usefulness in the Recovery of Protein-DNA Complexes from Avidin Affinity Columns”, Proc. Natl. Acad. Sci. USA, 1985, 82, 2593. |
6) W. J. LaRochelle and S. C. Froehner, “Immunodhemical Detection of Proteins Biotinylated on Nitrocellulose Replicas”, J. Immunol. Methods, 1986, 92, 65. |
7) P. S. Anjaneyulu and J. V. Staros,”Reactions of N-hydroxysulfosuccinimide Active Esters”, Int. J. Pept. Protein Res., 1987, 30, 117. |
8) H. M. Ingalls, C. M. Goodloe-Holland and E. J. Luna,”Junctional Plasma Membrane Domains Isolated from Aggregating Dictyostelium Discoideum Amebae”, Proc. Natl. Acad. Sci. USA, 1986, 83, 4779. |
9) J. Guesdon, T. Ternyck and S. Avrameas,”The Use of Avidin-Biotin Interaction in Immunoenzymatic Techniques”, J. Histochem. Cytochem., 1979, 27, 1131. |
上海金畔生物科技有限公司代理日本同仁化学 DOJINDO代理商全线产品,欢迎访问官网了解更多信息
Sulfo-AC5-SPDP试剂 S359 IgG纯化分离
蛋白抗体标记
品名货号用途
Sulfo-AC5-SPDP试剂 | S359 | IgG纯化分离 |
生化分析试剂 Biochemical Assay Reagents
Sulfo-SMCC sodium; 纯度: 97.24%
Sulfo-SMCC sodium 是一种常用的异双官能交联剂,是不可降解的 ADC linker,其带有N-羟基琥珀酰亚胺 (NHS) 酯和马来酰亚胺基团,分别与伯胺和巯基反应。
Sulfo-SMCC sodium Chemical Structure
CAS No. : 92921-24-9
规格 | 价格 | 是否有货 | 数量 |
---|---|---|---|
10 mg | ¥500 | In-stock | |
50 mg | ¥1300 | In-stock | |
100 mg | ¥2300 | In-stock | |
200 mg | ; | 询价 | ; |
500 mg | ; | 询价 | ; |
* Please select Quantity before adding items.
生物活性 |
Sulfo-SMCC sodium is a commonly used hetero-bifunctional, noncleavable ADC crosslinker bearing N-hydroxysuccinimide (NHS) ester and maleimide groups to react with primary amines and sulfhydryl groups, respectively. |
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IC50 Target |
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体外研究 (In Vitro) |
The crosslinker Sulfo-SMCC consists of a maleimide and an N-hydroxysuccinimide ester group to bind to sulfhydryl groups and primary amines, respectively. Sulfo-SMCC inhibits the end-to-end annealing of stabilized Microtubules (MTs). MTs are treated with 250 μM Sulfo-SMCC, and imaged after incubation for 0 h, 6 h, and 24 h. MTs treated with Sulfo-SMCC shows a constant mean length, independent of the incubation time[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only. |
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分子量 |
436.37 |
||||||||||||||||
Formula |
C16H17N2NaO9S |
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CAS 号 |
92921-24-9 |
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运输条件 |
Room temperature in continental US; may vary elsewhere. |
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储存方式 |
4deg;C, sealed storage, away from moisture and light *In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture and light) |
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溶解性数据 |
In Vitro:;
DMSO : 125 mg/mL (286.45 mM; Need ultrasonic) H2O : 5 mg/mL (11.46 mM; ultrasonic and warming and heat to 60°C) 配制储备液
*
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 In Vivo:
请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液,再依次添加助溶剂: ——为保证实验结果的可靠性,澄清的储备液可以根据储存条件,适当保存;体内实验的工作液,建议您现用现配,当天使用; 以下溶剂前显示的百
|
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参考文献 |
|
Kinase Assay [1] |
Labeled and unlabeled porcine tubulin powders are used. Sulfo-SMCC is used. MTs are polymerized to a final concentration of 2 mg/mL in 80 mM PIPES buffer (pH 6.8) containing 10 μM taxol, 2 mM MgCl2, 0.5 mM EGTA, and 1 mM GTP. A mixture of rhodamine-labeled tubulin and unlabeled tubulin (1:5) is used in the time-dependent measurements. In these experiments, polymerized MTs are divided in 2 groups: control and Sulfo-SMCC (250 μM) treated. Both samples are imaged after diluting 1:5 and incubating for 0 h (within 45 minutes after polymerization), 6 h, and 24 h[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only. |
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参考文献 |
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荧光染料Dye Reagents Cy5-DBCO;(Synonyms: DBCO-Sulfo-Cy5)
Cy5-DBCO (DBCO-Sulfo-Cy5) 是一个近红外 (NIR) 红色荧光染料 λabs 和 λem 值分别为 646 nm 和 670 nm。Cy5-DBCO (DBCO-Sulfo-Cy5) 不适合对通透性细胞的胞内成分进行染色,可能会表现出较高的背景。
Cy5-DBCO Chemical Structure
CAS No. : 1564286-24-3
规格 | 价格 | 是否有货 | |
---|---|---|---|
1 mg | ¥6500 | 询问价格 货期 | |
5 mg | ¥14500 | 询问价格 货期 |
* Please select Quantity before adding items.
生物活性 |
Cy5-DBCO (DBCO-Sulfo-Cy5) is a near-infrared (NIR) red fluorescent dye with λabs and λem of 646 nm and 670 nm, respectively. Cy5-DBCO (DBCO-Sulfo-Cy5) is not suitable for staining intracellular components of permeabilezed cell, it may exhibits a high background. |
IC50 Target |
Red fluorescent dye |
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体外研究 (In Vitro) |
Cy5-DBCO (DBCO-Sulfo-Cy5) reacts with azides via a copper-free “click chemistry” reaction and forms a stable triazole and does not need Cu-catalyst or elevated temperatures. MCE has not independently confirmed the accuracy of these methods. They are for reference only. |
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分子量 |
1009.22 |
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Formula |
C52H56N4O11S3 |
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CAS 号 |
1564286-24-3 |
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运输条件 |
Room temperature in continental US; may vary elsewhere. |
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储存方式 |
|
荧光染料Dye Reagents Cy5-DBCO;(Synonyms: DBCO-Sulfo-Cy5)
Cy5-DBCO (DBCO-Sulfo-Cy5) 是一个近红外 (NIR) 红色荧光染料 λabs 和 λem 值分别为 646 nm 和 670 nm。Cy5-DBCO (DBCO-Sulfo-Cy5) 不适合对通透性细胞的胞内成分进行染色,可能会表现出较高的背景。
Cy5-DBCO Chemical Structure
CAS No. : 1564286-24-3
规格 | 价格 | 是否有货 | |
---|---|---|---|
1 mg | ¥6500 | 询问价格 货期 | |
5 mg | ¥14500 | 询问价格 货期 |
* Please select Quantity before adding items.
生物活性 |
Cy5-DBCO (DBCO-Sulfo-Cy5) is a near-infrared (NIR) red fluorescent dye with λabs and λem of 646 nm and 670 nm, respectively. Cy5-DBCO (DBCO-Sulfo-Cy5) is not suitable for staining intracellular components of permeabilezed cell, it may exhibits a high background. |
IC50 Target |
Red fluorescent dye |
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---|---|---|---|---|---|---|---|---|---|---|
体外研究 (In Vitro) |
Cy5-DBCO (DBCO-Sulfo-Cy5) reacts with azides via a copper-free “click chemistry” reaction and forms a stable triazole and does not need Cu-catalyst or elevated temperatures. MCE has not independently confirmed the accuracy of these methods. They are for reference only. |
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分子量 |
1009.22 |
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Formula |
C52H56N4O11S3 |
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CAS 号 |
1564286-24-3 |
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运输条件 |
Room temperature in continental US; may vary elsewhere. |
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储存方式 |
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CY5;(Synonyms: Sulfo-Cyanine5) 纯度: 99.66%
Cy5 (Sulfo-Cyanine5) 是一种用于标记肽, 蛋白质, 寡核苷酸的氨基基团的反应染料。 激发波长 (nm): 649, 发射波长 (nm): 670。
CY5 Chemical Structure
CAS No. : 146368-11-8
规格 | 价格 | 是否有货 | 数量 |
---|---|---|---|
10;mM;*;1 mL in DMSO | ¥1878 | In-stock | |
5 mg | ¥1300 | In-stock | |
10 mg | ¥2000 | In-stock | |
25 mg | ¥4200 | In-stock | |
50 mg | ¥6700 | In-stock | |
100 mg | ; | 询价 | ; |
200 mg | ; | 询价 | ; |
* Please select Quantity before adding items.
生物活性 |
Cy5 (Sulfo-Cyanine5) is a reactive dye for the labeling of amino-groups in peptides, proteins, and oligonucleotides. This dye requires small amount of organic co-solvent (such as DMF or DMSO) to be used in labeling reaction. This reagent is ideal for very cost-efficient labeling of soluble proteins, as well as all kinds of peptides and oligonucleotides. This reagent also works well in organic solvents for small molecule labeling. Excitation (nm):649, Emission (nm): 670. |
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分子量 |
656.81 |
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Formula |
C33H40N2O8S2 |
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CAS 号 |
146368-11-8 |
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运输条件 |
Room temperature in continental US; may vary elsewhere. |
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储存方式 |
4deg;C, protect from light *In solvent : -80deg;C, 6 months; -20deg;C, 1 month (protect from light) |
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溶解性数据 |
In Vitro:;
DMSO : 250 mg/mL (380.63 mM; Need ultrasonic) H2O : 25 mg/mL (38.06 mM; Need ultrasonic) 配制储备液
*
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 In Vivo:
请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液,再依次添加助溶剂: ——为保证实验结果的可靠性,澄清的储备液可以根据储存条件,适当保存;体内实验的工作液,建议您现用现配,当天使用; 以下溶剂前显示的百
|
荧光染料Dye Reagents CY5-YNE;(Synonyms: Sulfo-Cyanine5-alkyne) 纯度: ge;95.0%
CY5-YNE (Sulfo-Cyanine5-alkyne) 是一种用于标记肽, 蛋白质, 寡核苷酸的氨基基团的反应染料。
CY5-YNE Chemical Structure
CAS No. : 1345823-20-2
规格 | 价格 | 是否有货 | 数量 |
---|---|---|---|
10;mM;*;1 mL in DMSO | ¥5343 | In-stock | |
5 mg | ¥3500 | In-stock | |
10 mg | ¥5600 | In-stock | |
25 mg | ¥10500 | In-stock | |
50 mg | ; | 询价 | ; |
100 mg | ; | 询价 | ; |
* Please select Quantity before adding items.
生物活性 |
CY5-YNE (Sulfo-Cyanine5-alkyne) is a reactive dye for the labeling of amino-groups in peptides, proteins, and oligonucleotides. |
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分子量 |
693.87 |
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Formula |
C36H43N3O7S2 |
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CAS 号 |
1345823-20-2 |
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运输条件 |
Room temperature in continental US; may vary elsewhere. |
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储存方式 |
4deg;C, protect from light *In solvent : -80deg;C, 6 months; -20deg;C, 1 month (protect from light) |
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溶解性数据 |
In Vitro:;
DMSO : ≥ 43 mg/mL (61.97 mM) * “≥” means soluble, but saturation unknown. 配制储备液
*
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 |