ELISA TMB Substrate for HRP

Cat. No.
250 ml
Cat. No. G443
Name ELISA TMB Substrate for HRP
Unit 250 ml
Category Gels, Blots & ELISAs

3,3'5,5'-Tetramethylbenzidine (TMB) is a safe-sensitive substrate used for the detection of Horseradish Peroxidase (HRP) activity. It yields a blue colour (Amax = 652nm) and changes to yellow (Amax = 450nm) upon the addition of either sulfuric or phosphoric acid. ABM’s ELISA TMB Substrate for HRP is a highly sensitive single component reagent that is ready to use for the quantitative detection of HRP bound to a solid phase or in free solution.

Concentration N/A

Suitable for kinetic and endpoint assays involving Horseradish Peroxidase (HRP).

Recommended Protocol 1. Complete all required incubations with antibodies, probes and HRP labeled reagents.
2. Wash plate, at least 4 times, with phosphate buffered saline or Tris buffered saline containing 0.1% Tween-20.
3. After the final wash, shake and blot all residual buffers from plate wells.
4. Add 0.1ml of TMB Substrate for HRP to the appropriate wells and incubate 5-30 minutes.
5. (Optional Higher Sensitivity Reaction Stop) Highest sensitivity of detection is achieved by stopping the TMBHK reaction with an equal volume of 0.1ml of 0.3M of sulphuric acid. Read the more highly absorbing yellow chromogen in the stopped reaction at 450nm.
6. Assay kinetics may be monitored at 650nm as a function of time. The reaction may be stopped to preserve the blue chromogen using an equal volume of 0.1ml of 0.1% sodium fluoride or 0.15% sodium dodecyl sulfate. Measure the blue chromogen in the stopped reaction at 650nm.
Storage Condition

Store at 2-4°C and then warm up to assay temperature prior to use. ELISA TMB Substrate for HRP can also be stored at room temperature for up to 6 months. Prevent exposure to direct sunlight. Discard if solution turns blue or turbid.


abm guarantees that all our Tissue Slides will perform as described on this product webpage, if this is not the case we will provide you with a one-time replacement at no extra cost. Documentation and explanation of experiment conducted will be required when submitting a claim for replacement.

Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G443
Print & Download Datasheet
Search CoA here
There are no Documents for this product yet!
There are no FAQs for this product yet!
  • Löffler, T et al. "Decreased Plasma Aβ in Hyperlipidemic APPSL Transgenic Mice Is Associated with BBB Dysfunction" Frontiers in Neuroscience 232 (10): (2016). DOI: 10.3389/fnins.2016.00232. Application: ELISA.
This product has no review yet.
Controls and Related Product:

500 μl/100 loads

100 μg

100 μg

100 μg