Alert® K-Blue® Aqueous is specifically designed for use with horseradish peroxidase (HRP) based sandwich immunoassays and also has been proven to work in competitive peroxidase-based immunoassays. The substrate is not intended for use with assays requiring a precipitating substrate. Alert K-Blue Aqueous utilises an indicator system that clearly indicates when substrate and acid stop have been added to an assay’s microwells.
The substrate is based on Neogen’s original K-Blue Aqueous formula which is a revolutionary 100% solvent free, non-volatile TMB substrate. It contains 3,3’,5,5’ tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2) in a one-bottle format with long-term stability and high activity. The addition of acid to the substrate produces a color change that can be measured on a microplate reader or observed visually. This color change increases the absorbance value at 540nm and changes the appearance of the substrate colors to pink.
Sensitivity: This is highly sensitive TMB substrate has low background development and offers excellent signal-to-noise ratio.
Stability and Storage: Alert K-Blue Aqueous is stable for 36 months when stored at 4°C.
Appearance: Yellow to golden yellow solution.
Custom Packaging Service: Neogen can package Alert K-Blue Aqueous in custom bottle sizes and volume fills to meet your specific packaging requirements. This service is a time and cost saving feature for any test kit manufacturer. For details on this service, please contact a Neogen representative.
Order Details
| Product # | Product Description |
| 334175 | Alert K-Blue Aqueous TMB Substrate, 200 mL |
| 334176 | Alert K-Blue Aqueous TMB Substrate, 500 mL |
| 334177 | Alert K-Blue Aqueous TMB Substrate, 1 L |
| 334257 | Alert K-Blue Aqueous TMB Substrate, 20 L (1 x 20 L) |
For research and manufacturing use only.
Individual well management is improved by substituting a Neogen Alert substrate for the traditional TMB substrate because the chance of a well returning a false negative due to reagent addition errors is reduced. This substrate change allows for an additional wavelength check to ensure that the only valid results are from wells that return a signal (pink) at 540 nm. As the figure below demonstrates, non-verifiable and false negatives with a traditional TMB substrate are recorded as invalid results when using the Alert TMB system.
| Traditional TMB Substrate with Acid Stop OD Measured at 450 nM - 650 nM | |||||
|---|---|---|---|---|---|
| Reagent Added: | Well contains sample that is: | ||||
| Substrate | Acid | NEGATIVE | Result | POSITIVE | Result |
| YES | YES | Colorless | TN | Yellow | TP |
| YES | NO | Colorless | N | Blue | Invalid or FN |
| NO | YES | Colorless | N | Colorless | FN |
| NO | NO | Colorless | N | Colorless | FN |
| Alert TMB Substrate with Acid Stop OD Measured at 450 nM - 650 nM | |||||
|---|---|---|---|---|---|
| Reagent Added: | Well contains sample that is: | ||||
| Substrate | Acid | NEGATIVE | Result | POSITIVE | Result |
| YES | YES | Colorless/No Pink | TN | Yellow/Pink | TP |
| YES | NO | Colorless/No Pink | Invalid | Blue/No Pink | Invalid |
| NO | YES | Colorless/No Pink | Invalid | Colorless/No Pink | Invalid |
| NO | NO | Colorless/No Pink | Invalid | Colorless/No Pink | Invalid |
TN = True Negative
N = Negative, but not verifiable
TP = True Positive
FN = False Negative
Invalid = Value returned in outside acceptable parameters