The Aladdin series of TMB ELISA substrates features low background, excellent lot-to-lot consistency, high stability, and multiple sensitivity options—providing a superior signal development system for ELISA kit R&D and manufacturing. Single-Component TMB ELISA Substrate HD is an ultra-sensitive TMB substrate that generates color rapidly and efficiently while maintaining low background and stable performance. It is recommended for assays with a 2–10 minute color development window.

Catalog No.: T615600-SS, HS, ST, BR, SL, SP

Pack sizes: 125 mL / 125 mL / 500 mL

pH: 3.6 ± 0.4

Storage: 2–8 °C

Application: ELISA

Instructions for Use

Important: Before use, bring Aladdin Single-Component TMB ELISA Substrate HD to the intended reaction temperature.

Endpoint read at 450 nm

1. Wash：After incubation with the HRP conjugate, wash the plate 3× with PBST/TBST washing buffer (with surfactant; Cat. E616323).

2. Develop：Add 100 µL substrate to each well and incubate at 20–37 °C for the desired time. The reaction develops a blue color.

3. Stop：Add 100 µL TMB Stop Solution 450 to each well. The color changes to yellow.

4. Read：Measure absorbance at 450 nm. Reading within 30 minutes is recommended.

Endpoint read at 650 nm

1. Wash：After incubation with the HRP conjugate, wash the plate 3× with PBST/TBST washing buffer (with surfactant; Cat. E616323).

2. Develop：Add 100 µL substrate to each well and incubate at 20–37 °C for the desired time. The reaction develops a blue color.

3. Stop：Add 100 µL TMB Stop Solution 650 to each well. The color remains blue.

4. Read：Measure absorbance at 650 nm. Reading within 30 minutes is recommended.

Kinetic read at 650 nm

1. Wash：After incubation with the HRP conjugate, wash the plate 3× with PBST/TBST washing buffer (with surfactant; Cat. E616323).

2. Develop：Add 100 µL substrate to each well and incubate at 20–37 °C.

3. Read：Record absorbance at 650 nm at the predetermined frequency (e.g., every 1 minute).

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Notice

For research use only or for further manufacture of in vitro diagnostic kits. Not for use in humans or animals.

Aladdin: https://www.aladdinsci.com/