Substrate Reagent Pack

R&D Systems | Catalog # DY999

New Version Available: DY999B
NEW
8 vials Color A, 8 vials Color B
R&D Systems
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Key Product Details

Applications

ELISA

Product Summary for Substrate Reagent Pack

INTENDED USE & DESCRIPTION

Substrate Reagent Pack contains 8 vials of Color Reagent A (stabilized hydrogen peroxide, Part # 895000) and 8 vials of Color Reagent B (stabilized tetramethylbenzidine, Part # 895001), both at 12 mL/vial, producing color development when contacting HRP-conjugated or biotinylated, streptavidin-HRP bound detection antibodies.   

STORAGE & STABILITY

Store unopened at 2-8 °C. Stable for up to 30 days after opening when stored at 2-8 °C.

REAGENT PREPARATION

Warm to room temperature before use. Color Reagent A and Color Reagent B should be mixed together in equal volumes within 15 minutes of use. Protect from light. Add 100 µL of the mixture to each well and incubate at room temperature for 20 minutes while protecting from light. Add Stop Solution (Catalog # DY994) and read the microplate.

TECHNICAL HINTS & LIMITATIONS OF THE PROCEDURE 

  • Caution: Color Reagent B may cause skin, eye, and respiratory irritation. Avoid breathing fumes. Wear protective gloves, clothing, and eye and face protection. Wash hands thoroughly after handling. Please refer to the MSDS on our website prior to use. 

Formulation, Preparation, and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Product Documents for Substrate Reagent Pack

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Substrate Reagent Pack

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Substrate Reagent Pack

Customer Reviews for Substrate Reagent Pack (54)

4.8 out of 5
54 Customer Ratings
5 Stars
78%
4 Stars
20%
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1 Stars
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  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 01/02/2026
    Used in Angiogenin Duoset ELISA to test conditioned media of adipose tissue and coculture of HCT116 and adipose tissue
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 01/02/2026
    SVF conditioned media analysed wit IL-8 ELISA Duoset
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Maximiliaan Hennink
    Verified Customer | Posted 08/06/2025
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 05/25/2024
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 05/21/2024
    Works well for Duoset kits
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 06/23/2023
  • Substrate Reagent Pack
    Name: Do-Hyun Kim
    Verified Customer | Posted 03/28/2023
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 02/27/2023
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 05/23/2022
    Substrate works well in direct and sandwich ELISAs
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 05/13/2021
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 01/12/2021
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 12/17/2020
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Gagandeep Kaur
    Verified Customer | Posted 11/11/2020
  • Substrate Reagent Pack
    Name: Walker McHugh
    Verified Customer | Posted 10/23/2020
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 08/11/2020
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Kumudhini Haran
    Verified Customer | Posted 07/13/2020
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 06/20/2020
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 02/11/2020
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 01/17/2020
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 01/10/2020
  • Substrate Reagent Pack
    Name: Tahmineh Safaie
    Verified Customer | Posted 12/23/2019
    One pair of A and B is enough for one plate which makes it very easy to use while keep the reagent stable
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 09/07/2019
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 07/02/2019
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 03/28/2019
    I used this a my detection substrate (100uL) in a direct ELISA after my plate was treated with an anti-mouse IgG-HRP antibody. I find the background is very low which allows me to achieved a better sensitivity with my ELISAs compared another commercial substrate
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 03/13/2019
  • Substrate Reagent Pack
    Name: Ivan Luzardo
    Verified Customer | Posted 01/03/2019
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 12/10/2018
  • Substrate Reagent Pack
    Name: Marry Tahani
    Verified Customer | Posted 11/21/2018
    It is very stable before mixing the reagent A and B. One bottle of each is exactly enough for two plates which makes measuring simple.
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Sijia He
    Verified Customer | Posted 11/13/2018
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 09/30/2018
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 08/17/2018
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 06/19/2018
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 06/15/2018
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 05/10/2018
  • Substrate Reagent Pack
    Name: Leslie Priddy
    Verified Customer | Posted 03/30/2018
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 02/22/2018
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 02/06/2018
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 02/06/2018
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack
    Name: Anonymous
    Verified Customer | Posted 01/19/2018
  • Substrate Reagent Pack
    Name: Balaji Mahender
    Verified Customer | Posted 12/20/2017
  • Substrate Reagent Pack
    Name: Jomkuan Theprungsirikul
    Verified Customer | Posted 12/15/2017
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Jomkuan Theprungsirikul
    Verified Customer | Posted 12/05/2017
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: David Bolick
    Verified Customer | Posted 09/07/2017
    Our lab does many ELISAs. These substrate solutions are excellent, affordable, and easy to use.
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Anonymous
    Verified Customer | Posted 07/19/2017
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Anonymous
    Verified Customer | Posted 07/17/2017
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Anonymous
    Verified Customer | Posted 07/14/2017
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Anonymous
    Verified Customer | Posted 07/06/2017
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Goutham Pattabiraman
    Verified Customer | Posted 06/15/2017
    Really Easy to use. After the SA-HRP detection step for ELISA, wash wells 4 times with wash buffer, and add the Substrate and incubate for 20 mins in the dark. Stop the reaction using 0.2 M sulfuric acid. Read the plate at 450nm
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Abby Sukarto
    Verified Customer | Posted 05/03/2017
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Anonymous
    Verified Customer | Posted 03/29/2017
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Anonymous
    Verified Customer | Posted 11/28/2016
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Adam Guess
    Verified Customer | Posted 09/01/2016
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: John Yu
    Verified Customer | Posted 04/26/2016
    Substrate Reagent Pack DY999
  • Substrate Reagent Pack (8 vials Color A, 8 vials Color B)
    Name: Anonymous
    Verified Customer | Posted 01/18/2016

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FAQs for Substrate Reagent Pack

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    • A: Yes, TMB ELISA Substrate (Catalog # DY999B) has been tested for equivalency and can be substituted for a 1:1 mixture of Color Reagent A and B (Catalog # DY999).
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