Alert® K-Blue® Aqueous TMB Substrate

Item No.  334175

  • 100% solvent-free
  • Clearly indicates when substrate and acid stop have been added to the wells (turns pink)
  • Ideal for sandwich based assays
  • Low background
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$94.40
$132.00
$236.20
$3,070.20
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Minimum quantity required: 1
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Alert® K-Blue® Aqueous utilizes 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 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 color to pink.
Specifications
Application Microwell
Brand K-Blue®
Enzyme HRP
Intended Use Research and manufacturing use only
Result Type Colorimetric
Sensitivity High Activity
Storage Conditions Stable for 36 months when stored at 2-8°C.
Wavelength 540 nm

Additional Information

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


Training

Our customers’ success is our shared success. Our experts are ready to train you and your team on our solutions, so you can rest easy knowing procedures are performed properly and yield accurate results. In addition, we provide certificates upon completion of training.