Antibody Sensitizer (IC-8012)

Introduction

IC-8012 Antibody Sensitizer is a proprietary, polymer-based reagent designed to significantly amplify the signal in immunohistochemistry (IHC) and immunocytochemistry (ICC) applications. It works by enhancing the binding of horseradish peroxidase (HRP)-conjugated secondary antibodies and subsequently increasing the deposition of chromogen, leading to intense, high-contrast staining without increasing non-specific background. This allows for the detection of low-abundance antigens and enables the use of higher antibody dilutions, reducing costs.

Specifications

-Appearance: Clear, colorless to slightly yellowish liquid.

-Formulation: Aqueous buffer solution with proprietary polymers and stabilizers.

-Concentration: Ready-to-use solution.

-Storage: Store at 2¡ãC to 8¡ãC. Do not freeze.

-Stability: Stable for 24 months from the date of manufacture when stored properly.

-Compatibility: Compatible with HRP-based detection systems and common chromogens like DAB (3,3''-Diaminobenzidine).

Principle of the Assay

The sensitizer contains multiple branched polymer chains coated with secondary antibody molecules (e.g., anti-mouse/anti-rabbit IgG) and a high number of HRP enzymes. This structure allows for the following mechanism:

1.  The polymer-bound secondary antibodies bind specifically to the primary antibody.

2.  Each polymer molecule carries dozens of HRP enzymes to the antigen site.

3.  During the chromogenic reaction, this massive localized enzyme load catalyzes a robust deposition of precipitate (e.g., DAB), resulting in a dramatically amplified signal compared to a standard secondary antibody.

Kit Components

Quantity: 15 ml/ 50 ml / 100 ml /500ml

Protocol (General Guidelines)

Important: Optimize incubation times, temperatures, and antibody dilutions for your specific tissue and primary antibody.

A. Two-Step Protocol (Recommended for most applications)

1.  Deparaffinization, Antigen Retrieval, and Blocking: Perform standard procedures on your tissue sections.

2.  Primary Antibody: Apply appropriately diluted primary antibody. Incubate, then wash.

3.  Sensitizer Application: Apply ready-to-use IC-8012 Antibody Sensitizer to cover the tissue section. Incubate for 15-30 minutes at room temperature.

4.  Wash: Rinse thoroughly with PBS or TBS buffer (3 x 5 minutes).

5.  Chromogen Detection: Apply your preferred HRP substrate (e.g., DAB) and monitor development.

6.  Counterstain, Dehydrate, and Mount.

B. One-Step Protocol (Primary antibody must be from mouse or rabbit)

1.  Prepare Working Solution: Mix your mouse or rabbit primary antibody directly with the IC-8012 Sensitizer at the optimal ratio (e.g., 1:50 to 1:200 of primary antibody in sensitizer). This mixture must be prepared immediately before use.

2.  Apply the antibody-sensitizer mixture to the prepared tissue section.

3.  Incubate for 60-120 minutes at room temperature.

4.  Wash and proceed with chromogen detection as in steps 4-6 above.

Safety and Handling

Precaution: This product contains material of biological origin. Handle as though capable of transmitting infectious agents.

Personal Protective Equipment: Wear lab coat, gloves, and safety glasses.

Disposal: Dispose of waste in accordance with all applicable local, state, and federal regulations.

Intended Use: Research Use Only (RUO). Not for use in diagnostic procedures.

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