DTNB(Synonyms: Ellman’s Reagent)

生化分析试剂 Biochemical Assay Reagents
DTNB;(Synonyms: Ellman’s Reagent) 纯度: 98.35%

DTNB (Ellman’s Reagent) 是一种用于定量巯基数量或浓度的化学物质。常用于检测游离巯基。

DTNB(Synonyms: Ellman’s Reagent)

DTNB Chemical Structure

CAS No. : 69-78-3

规格 价格 是否有货 数量
10;mM;*;1 mL in DMSO ¥500 In-stock
500 mg ¥350 In-stock
1 g ¥450 In-stock
5 g ¥650 In-stock
10 g ; 询价 ;
50 g ; 询价 ;

* Please select Quantity before adding items.

DTNB 相关产品

bull;相关化合物库:

  • Covalent Screening Library Plus
  • Bioactive Compound Library Plus
  • Covalent Screening Library

生物活性

DTNB (Ellman’s Reagent) is a chemical used to quantify the number or concentration of thiol groups[1].

体外研究
(In Vitro)

DTNB reacts with the free sulfhydryl side chain of cysteine to form an S-S bond between the protein and a thionitrobenzoic acid (TNB) residue[1].
The main advantage of DTNB over alternative reagents (e.g., N-ethylmaleimide or iodoacetamide) is in the selectivity of this reagent and in the ability to follow the course of the reaction spectrophotometrically[1].
Modification of Enzyme with DTNB:
Modification of the SH groups of the enzyme is carried out by reacting 10 μL of 10 mM DTNB solution (about a 20-fold molar excess) at room temperature with 0.6 mL of enzyme solution (0.807 mg/mL) in 80 mM phosphate buffer, pH 8.0, containing 1 mM EDTA, which has been dialyzed previously against the same buffer solution for 24 h. The number of SH groups is estimated from the increase of absorbance at 410 nm using a molar extinction coefficient of 13,600 M-1cm-1 for thionitrobenzoate ions liberated.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

分子量

396.35

Formula

C14H8N2O8S2

CAS 号

69-78-3

运输条件

Room temperature in continental US; may vary elsewhere.

储存方式
Powder -20deg;C 3 years
4deg;C 2 years
In solvent -80deg;C 6 months
-20deg;C 1 month
溶解性数据
In Vitro:;

DMSO : ≥ 150 mg/mL (378.45 mM)

* “≥” means soluble, but saturation unknown.

配制储备液
浓度 溶剂体积 质量 1 mg 5 mg 10 mg
1 mM 2.5230 mL 12.6151 mL 25.2302 mL
5 mM 0.5046 mL 2.5230 mL 5.0460 mL
10 mM 0.2523 mL 1.2615 mL 2.5230 mL

*

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
储备液的保存方式和期限:-80°C, 6 months; -20°C, 1 month。-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。

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
分比是指该溶剂在您配制终溶液中的体积占比;如在配制过程中出现沉淀、析出现象,可以通过加热和/或超声的方式助溶

  • 1.

    请依序添加每种溶剂:;10% DMSO ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

    Solubility: ≥ 2.08 mg/mL (5.25 mM); Clear solution

    此方案可获得 ≥ 2.08 mg/mL (5.25 mM,饱和度未知) 的澄清溶液。

    以 1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中,混合均匀;向上述体系中加入50 μL Tween-80,混合均匀;然后继续加入 450 μL生理盐水定容至 1 mL。

    将 0.9 g 氯化钠,完全溶解于 100 mL ddH₂O 中,得到澄清透明的生理盐水溶液

  • 2.

    请依序添加每种溶剂:;10% DMSO ;; 90% (20% SBE-β-CD in saline)

    Solubility: ≥ 2.08 mg/mL (5.25 mM); Clear solution

    此方案可获得 ≥ 2.08 mg/mL (5.25 mM,饱和度未知) 的澄清溶液。

    以 1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中,混合均匀。

    将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中,再用生理盐水定容至 10 mL,完全溶解,澄清透明
  • 3.

    请依序添加每种溶剂:;10% DMSO ;; 90% corn oil

    Solubility: ≥ 2.08 mg/mL (5.25 mM); Clear solution

    此方案可获得 ≥ 2.08 mg/mL (5.25 mM,饱和度未知) 的澄清溶液,此方案不适用于实验周期在半个月以上的实验。

    以 1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中,混合均匀。

*以上所有助溶剂都可在 MCE 网站选购。
参考文献
  • [1]. T Nagaoka, et al. DTNB modification of SH groups of isocitrate dehydrogenase from Bacillus stearothermophilus purified by affinity chromatography. J Biochem. 1977 Jan;81(1):71-8.