TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)

製品の概要

  • 製品名

    TMB ELISA Substrate (Fast Kinetic Rate)
    ELISA substrate ihc 試薬 製品一覧
  • アプリケーション

    適用あり: ELISAmore details
    適用なし: IHC-Fr or IHC-P
  • 特記事項

    Abcam’s TMB ELISA Substrate (Fast Kinetic Rate) detects horseradish peroxidase (HRP) activity and contains 3,3’,5,5’-tetramethylbenzidine in a mildly acidic buffer. The substrate is supplied as a ready to use solution. Unreacted substrate should be colorless or very light yellow in appearance. When this substrate system is reacted with peroxidase, a soluble blue reaction product is obtained. The reaction can be stopped using appropriate stop solution, producing a soluble yellow or soluble blue reaction product, depending upon the stop reagent used, which is stable for at least 1 hour. ab171524 is not recommended for membrane or immunohistochemical applications that require a precipitating reaction product.

    Product Use:
    ab171524 is supplied as a ready to use solution. The product should be allowed to equilibrate to room temperature (25°C) prior to use. For microwell applications, 100 µL of substrate solution is added to each well. A soluble blue reaction product develops which can be read at 370 nm or in a range of 620 nm to 650 nm. For best results, sample absorbance values should be monitored and read before absorbance values exceed 2.0 OD units. In endpoint assays, the substrate reaction can be stopped using equal volumes of 1 N HCl, 0.6 N sulfuric acid, or one of the stop solutions (ab171529 and ab171531). Addition of acid turns the blue color to yellow and stops the enzymatic reaction. In the case of the 650 nm Stop Solutions for TMB Substrate (ab171531), the blue color does not change. Since stopping the reaction increases sample absorbance values approximately three fold, unless using the 650 nm Stop Solutions for TMB Substrate (ab171531), sample OD values should be monitored and substrate reaction stopped when values reach approximately 0.7 OD units. After stopping with the 450 nm Stop Solutions for TMB Substrate (ab171529), a soluble yellow product develops which is read in the 450 nm range. Stopping with the 650 nm Stop Solutions for TMB Substrate (ab171531) produces a soluble blue product which is read in the 650 nm range. Dilution of the substrate is not recommended. To reduce the intensity of a reaction, it is recommended that the antibodies or conjugates be diluted.

    Storage Instructions:

    Product should be stored at 2-8°C. Exposure to direct sunlight and other UV sources should be avoided due to the light sensitive nature of the TMB molecule.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Store at +4°C short term (1-2 weeks). Store at +4°C. Please see notes section.
  • バッファー

    Constituents: 0.05% TMB, 20% Proprietary component, 79% Water
  • Concentration information loading...

アプリケーション

Our Abpromise guarantee covers the use of ab171524 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ELISA 1/1. 100 µL of substrate solution is added to each well.
  • 追加情報
    Is unsuitable for IHC-Fr or IHC-P.
  • 画像

    • Comparison of the kinetic curves of several TMB substrates at 500 pg/mL msIgG monitored over time at 650 nm.
    • Expanded graph at the lower concentrations using TMB ELISA Substrate (Fast Kinetic Rate) (ab171524) and TMB ELISA Substrate (Slowest Kinetic Rate) (ab171527). Both ab171524 and ab171527 have the same detection limit.
    • Comparison of the kinetic curves of TMB ELISA Substrate (Fast Kinetic Rate) (ab171524) at different concentrations of msIgG monitored over time at 650 nm.

    参考文献

    This product has been referenced in:

    See all 2 Publications for this product

    レビューと Q&A

    Answer



    ab171524 (TMB ELISA Substrate - Fast Kinetic Rate) and ab171527 (TMB ELISA Substrate - Slowest Kinetic Rate) differ in the rate of color development and sensitivity.

    TMB ELISA Substrate - Slowest Kinetic Rate has a relatively slower rate of color development and is a less sensitive substrate than ab171524 (TMB ELISA Substrate - Fast Kinetic Rate.

    In general, one would select ab171527 (TMB ELISA Substrate - Slowest Kinetic Rate) if you feel the target protein detected in your assay is present in high amounts. The slow color development will allow you to stop the assay in a reasonable amount of time (eg. 5-10 minutes).

    If you used ab171524 (TMB ELISA Substrate - Fast Kinetic Rate) as the substrate in the above case the color development may occur to quickly. This would lead to problems such as ODs quickly growing beyond the linear range of your spectrophotometer. In addition, while you are adding STOP solution to the first row of the plate, the remaining wells will continue to develop color while you add STOP solution to all the wells of the plate, leading to inconsistant results across the plate.

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    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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