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形状
liquid
メーカー／製品名
Chemicon®
テクニック
ELISA: suitable
λmax
450 nm
輸送温度
wet ice
詳細
TMB, (3,3′,5,5′-tetramethybenzidine) has been shown to be a safe-sensitive substrate for the assay of HRP. Initially, in the presence of HRP and hydrogen peroxide, a one-electron oxidation product is formed. This compound, a cation free radical, is blue in color with an adsorption maximum of 653 nm. Further reaction with HRP/H2O2 or acidification of the radical with acid yields the diimine terminal oxidation product adsorbing light at 450 nm. The extinction coefficient of the radical (E653nm=3.9 x 104 mol-1 cm-1) and diimine (E450nm=5.9 x 104 mol-1 cm-1) provide a remarkably sensitive system for the assay of HRP and HRP labeled probes. ES001 is a single component reagent stable at room temperature and not sensitive to normal laboratory light. It is optimized with respect to TMB and hydrogen peroxide concentrations and yields a linear response with the concentrations of HRP usually employed in immunological assays.
アプリケーション
METHOD SYNOPSIS:
After completion of analyte binding to a solid phase and reaction with a HRP labeled probe, ES001 is added. Oxidation of TMB produces a blue reaction product that is measured at 650 nm. The color formation as a function of time can be recorded or the reaction stopped with sodium fluoride after a fixed interval. Increased sensitivity can be achieved by converting the blue radical to the diimine by addition of acid. The resulting yellow color is measured immediately at 450 nm.
SAMPLE PROCEDURE
1. Complete all required incubations with antibodies, probes, HRP labeled reagents.
2. Wash plate at least 4 times with PBS or Tris buffered saline containing 0.1% Tween-20.
3. After the final wash, shake and blot all residual buffer from plate wells.
4. Add 0.1 mL of ES001 (TMBE Solution) to appropriate wells and incubate 5-30 minutes.
Note: The reaction time will depend upon the activity of the HRP probe. If color develops too briskly, zero order kinetics will not prevail. Dilution of the probe, antibody or HRP labeled reagent may be required.
5. The reaction can be monitored as a function of time for kinetic assays or stopped with 0.1 mL of 0.1% sodium fluoride and read at 650 nm.
6. If the procedure demands conversion to the yellow diimine, add 0.1 mL of either acid described in the reagent section above and record the absorbance within 5 minutes.
Variations of time, reagent volume and temperature require standardization by the user.
After completion of analyte binding to a solid phase and reaction with a HRP labeled probe, ES001 is added. Oxidation of TMB produces a blue reaction product that is measured at 650 nm. The color formation as a function of time can be recorded or the reaction stopped with sodium fluoride after a fixed interval. Increased sensitivity can be achieved by converting the blue radical to the diimine by addition of acid. The resulting yellow color is measured immediately at 450 nm.
SAMPLE PROCEDURE
1. Complete all required incubations with antibodies, probes, HRP labeled reagents.
2. Wash plate at least 4 times with PBS or Tris buffered saline containing 0.1% Tween-20.
3. After the final wash, shake and blot all residual buffer from plate wells.
4. Add 0.1 mL of ES001 (TMBE Solution) to appropriate wells and incubate 5-30 minutes.
Note: The reaction time will depend upon the activity of the HRP probe. If color develops too briskly, zero order kinetics will not prevail. Dilution of the probe, antibody or HRP labeled reagent may be required.
5. The reaction can be monitored as a function of time for kinetic assays or stopped with 0.1 mL of 0.1% sodium fluoride and read at 650 nm.
6. If the procedure demands conversion to the yellow diimine, add 0.1 mL of either acid described in the reagent section above and record the absorbance within 5 minutes.
Variations of time, reagent volume and temperature require standardization by the user.
物理的形状
Color Code: 黄
REAGENT PROVIDED:
Liquid solution containing TMB, 1.25 mMol/L and Hydrogen Peroxide, 2.21 mMol/L and less than 1% dimethyl sulfoxide in a 0.08 Mol/L) acetate buffer at pH 4.9. Also contains non-toxic proprietary stabilizers.
Materials Required:
Reagents Required for Stopping the Reaction, But Not Provided
A. Sodium fluoride, 0.1% for stopping the reaction and preserving blue chromogen. Prepare by dissolving 1 g of sodium fluoride in 1 liter of reagent grade water. Observe all precautions on label for use of sodium fluoride.
B. Acid for stopping the reaction and converting the blue radical to the yellow diimine. Either 0.5 Mol/L sulfuric acid or 0.25 Mol/L hydrochloric acid may be used. THE ABSORBANCE MUST BE OBTAINED WITHIN 5 MINUTES AFTER ADDING THE ACID.
Note: Reagent grade water must contain less than 10-7 Mol/L of iron or copper salts otherwise unreacted TMB will be converted non-enzymatically to the diimine.
Liquid solution containing TMB, 1.25 mMol/L and Hydrogen Peroxide, 2.21 mMol/L and less than 1% dimethyl sulfoxide in a 0.08 Mol/L) acetate buffer at pH 4.9. Also contains non-toxic proprietary stabilizers.
Materials Required:
Reagents Required for Stopping the Reaction, But Not Provided
A. Sodium fluoride, 0.1% for stopping the reaction and preserving blue chromogen. Prepare by dissolving 1 g of sodium fluoride in 1 liter of reagent grade water. Observe all precautions on label for use of sodium fluoride.
B. Acid for stopping the reaction and converting the blue radical to the yellow diimine. Either 0.5 Mol/L sulfuric acid or 0.25 Mol/L hydrochloric acid may be used. THE ABSORBANCE MUST BE OBTAINED WITHIN 5 MINUTES AFTER ADDING THE ACID.
Note: Reagent grade water must contain less than 10-7 Mol/L of iron or copper salts otherwise unreacted TMB will be converted non-enzymatically to the diimine.
保管および安定性
Maintain at room temperature for up to six months. For long term storage, up to twelve months, maintain at 2-8°C. Protect from exposure to direct sunlight. Discard if solution is blue or turbid.
法的情報
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責事項
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
保管分類コード
12 - Non Combustible Liquids
WGK
WGK 2
引火点(°F)
Not applicable
引火点(℃)
Not applicable
適用法令
試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。
労働安全衛生法名称等を通知すべき危険物及び有害物
名称等を通知すべき危険物及び有害物
Jan Code
ES001-100ML:
ES001-500ML:
ES001-1L-K:
試験成績書（COA）
製品のロット番号・バッチ番号を入力して、試験成績書（COA） を検索できます。ロット番号・バッチ番号は、製品ラベルに「Lot」または「Batch」に続いて記載されています。
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