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Tetramethylbenzidine (TMB) and assay buffers

We have blocking buffers and reagents to support your own immunoassay development including blocking, stability and detection reagents.

Additionally, we offer TMB (3,3’,5,5’-Tetramethylbenzidine), the most popular chromogenic substrate for HRP detection with different kinetic formulations to help optimize your particular application.

Blocking buffers and conjugate stabilizers

The buffers are specifically designed to preserve the conformational spatial arrangement of proteins in their native folded state when dried or in solution ensuring the biological activity of the protein.

Product name

Function

Product code

Immunoassay Blocking Buffer

Blocking agent that can be dried down or used for same-day assays. Preserves the conformation and activity of dried antibodies, proteins or enzymes coated to a surface.

ab171534

Immunoassay Blocking Buffer (BSA free)

Blocking solution without BSA. Synthetic formulation to eliminate background issues.

ab171535

AP Stability Conjugate Reagent

Preserves AP conjugated proteins in native folded state in solution

ab171536

HRP Stability Conjugate Reagent

Preserves HRP conjugated proteins in native folded state in solution

ab171537

Table 1: Buffers to preserve the conformational spatial arrangement of proteins

See below some comparative studies when using our buffers with monoclonal or polyclonal antibodies.

Figure 1: After 30 months in Immunoassay Blocking Buffer (ab171534) at room temperature, the monoclonal antibody retained 70% activity and the polyclonal antibody retained 95% activity.

Figure 2: After 30 months in Immunoassay Blocking and Protein Stabilizer (BSA free) (ab171535) at room temperature, the monoclonal antibody retained greater than 85% activity and the polyclonal antibody retained 90% activity.

Product name

Product code

Stabilization

ELISA

WB

Microspheres

Immunoassay Blocking Buffer

ab171534

ü

ü

ü

Immunoassay Blocking and Protein Stabilizer (BSA free)

ab171535

ü

ü

ü

ü

AP Stability Conjugate Reagent

ab171536

ü

RP Stability Conjugate Reagent

ab171537

ü

Table 2: Blocking and conjugate buffers guide

Different kinetic TMB formulations

The choice of the TMB substrate formulation optimizes the ELISA dynamic range as well as the assay sensitivity:

  • Detection of low concentration proteins of interest over a short period of time
  • Improved assay sensitivity and performance
  • Reliable standard curve

Product name

Function

Product code

TMB ELISA substrate (Highest Sensitive)

Most sensitive TMB formulation

ab171522

TMB ELISA substrate (High Sensitivity)

Formulated for liquid level detection systems on instruments

ab171523

TMB ELISA substrate (Fast Kinetic Rate)

Most popular and robust formulation

ab171524

TMB ELISA substrate (Slow Kinetic Rate)

Allow a wide dynamic/working range without compromising the assay sensitivity

ab171525

TMB ELISA substrate (Slower Kinetic Rate)

Option for maximum dynamic/working range

ab171526

TMB ELISA substrate (Slowest Kinetic Rate)

Option for extended dynamic range and improved reproducibility

ab171527

Table 3: Range of TMB substrates with different kinetic formulations

Figure 3: View the kinetics among different TMB formulations after a sandwich ELISA where the capture antibody was blocked with Immunoassay Blocking Buffer (ab171534) and the HRP conjugated detection antibody was diluted with HRP Stability Conjugate Reagent (ab171537).

SimpleStep ELISA kits
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