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Flagship Product: Safer, More Stable, and More Versatile – ProCide™ 300 Preservative!
Newly Upgraded 3rd-Gen TMB Substrate: Enhanced Stability with Multiple Freeze-Thaw Tolerance!
New Product Launch – Bovine Serum Albumin (BSA) for Clinical Biochemistry, Immunoassays, and Molecular Detection!
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TMB High Sensitive One Component ELISA Substrate
$165/ LELISA
Durastab™ TMB High Sensitive One Component ELISA Substrate is a ready-to-use, one-component chromogenic substrate for horseradish peroxidase (HRP) detection in ELISA. It oxidizes to a blue product measurable at 450 nm after stopping (or 650 nm without stopping), with a chromogenic time of 10–15 minutes at 20–37 °C. The low-background, high-stability formula is designed for the development and production of ELISA kits. Available in 100 mL, 1 L, 5 L, 10 L and bulk; store at 2–8 °C.
Product Parameters
- Art noTMHS
- Size100mL, 1L, 5L, 10L, bulk
- pH value3.6±0.4
- Shelf life3 year
- Storage2-8℃
- ApplicationELISA
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Description
TMB (3,3′,5,5′-tetramethylbenzidine) substrate is a chromogenic, colorimetric substrate used with horseradish peroxidase (HRP) in ELISA and other immunoassays.
In an ELISA, an HRP-labeled antibody is retained on the plate, and when TMB substrate solution is added, HRP oxidizes TMB in the presence of hydrogen peroxide, producing a blue soluble product whose intensity is proportional to the amount of analyte in the sample. After the reaction is stopped, the color shifts to yellow and is quantified at 450 nm.
Durastab™ TMB ELISA Substrate series features low background, high stability, minimal lot-to-lot variation, and options for different sensitivity levels. It provides an excellent signal response system suitable for the development and production of ELISA kits.TMB High Sensitive One Component ELISA Substrate has high sensitivity, low background and and reliable stability, which is suitable for detection assays with a chromogenic reaction time of 10–15 minutes.
Our TMB substrate is validated for sandwich, indirect, direct, and competitive ELISA, and for microwell HRP detection in quantitative assays.
Quality & Documentation
Every lot is released with a Certificate of Analysis (CoA) documenting QC criteria and batch-to-batch consistency.
Product Features & Benefits
• Ready-to-use and one-component – no mixing or dilution required, saving time and reducing error.
• High sensitivity and low background – reliable detection across a wide concentration range.
• Broad compatibility – works with HRP conjugates in sandwich, indirect, direct, and competitive ELISA.
• Stable and reproducible – consistent lot-to-lot performance with documented QC.
• Easy endpoint readout – blue at 370/650–652 nm, yellow at 450 nm after stop solution.
Instruction
Durastab™ TMB High Sensitive One Component ELISA Substrate should be equilibrated to room temperature before use.
ELISA Chromogenic Reaction Procedure：
Measurement at 450 nm:
1. Wash: After HRP conjugate incubation, wash the plate 3 times with PBST/TBST washing buffer (containing surfactant).
2. Chromogenic reaction: Add 100 μL substrate and incubate at 20–37 °C for the appropriate time, color is blue;
3. Stop: Add 100 μL TMB Stop Solution 450, the color will change yellow;
4. Read: Measure the absorbance at 450 nm within 30 minutes.
Measurement at 650nm：
1. Wash: After HRP conjugate incubation, wash the plate 3 times with PBST/TBST washing buffer (containing surfactant).
2. Chromogenic reaction: Add 100 μL substrate and incubate at 20–37 °C for the appropriate time, color is blue;
3. Stop: Add 100 μL TMB Stop Solution 650, the color remains blue ;
4. Read: Measure the absorbance at 650 nm within 30 minutes.
Kinetic measurement at 650 nm:
1. Wash: After HRP conjugate incubation, wash the plate 3 times with PBST/TBST washing buffer (containing surfactant).
2. Chromogenic reaction: Add 100 μL substrate and incubate at 20–37 °C.
3. Read: Measure the absorbance at 650 nm at set intervals (e.g., every 1 minute).
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Precautions
For research use only or for further use in the production of detection kits. Not for human or animal use!
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FAQ
Q: What is this TMB substrate used for?
A: This one-component TMB substrate is used with HRP in ELISA for chromogenic, colorimetric detection. It produces a blue color read at 450 nm (or 650 nm without stopping) and is suited to ELISA kit development and production.
Q: What wavelength should I read at?
A: Add stop solution and read at 450 nm within 30 minutes, or read the blue form at 650 nm without stopping. For kinetic monitoring, read at 650 nm at set intervals.
Q: How long does the chromogenic reaction take?
A: For the High Sensitive formulation, color develops over approximately 10–15 minutes at 20–37 °C. Optimize incubation toward your assay’s target signal and always follow the product instructions.
Q: How should I store it?
A: Store at 2–8 °C, protect from light, and bring to room temperature before use. Shelf life is 3 years under recommended storage.
Q: Is it for human or animal use?
A: No. It is for research use only or for further use in the production of detection kits. Not for human or animal use.
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