The MicroStacker™ Polymer Detection System Universal is a biotin-free, polymer-based horseradish peroxidase (HRP) secondary antibody detection system designed for use with mouse and rabbit primary antibodies in immunohistochemistry (IHC) on formalin-fixed, paraffin-embedded (FFPE) tissue samples.
Its proprietary MicroStacker™ technology enables precise, layered assembly of antibodies and HRP enzymes onto a micro-polymer scaffold. This creates a compact polymer structure with excellent tissue penetration, allowing access to all cellular compartments and delivering enhanced sensitivity compared with traditional HRP polymers that rely on bulky dextran backbones.
In addition, the system incorporates F(ab’)₂ fragments of IgG secondary antibodies rather than intact IgG molecules, minimizing nonspecific background staining caused by interactions with endogenous Fc receptors.
Recommended Protocol
Deparaffination: Deparaffination slides in xylene. Hydrate slides in a series of graded alcohols to water.
Peroxidase Block: Block for 5mins with Peroxidase Blocking Reagents. Rinse 2 times with wash buffer for 5mins each.
Antigen Retrieval Protocol: Please refer to the respective primary antibody data sheet for recommended antigen retrieval protocol.
Primary Antibody: Apply 2 drops (100ul) or as much as needed of ready-to-use primary antibody to cover the specimen. Incubate for 30-60mins at RT. Rinse 2 times with wash buffer for 5mins each.
Detection Polymer: Apply 2 drops (100ul) or as much as needed of anti-Mouse/Rabbit HRP-Polymer to cover the specimen. Incubate for 30mins at RT. Rinse 2 times with wash buffer for 5mins each.
DAB Chromogen: Prepare ready-to-use DAB substrate solution. Add DAB Chromogen to DAB Substrate Buffer and mix the two solutions in a 1:20 ratio with the volume determined by the number of slides to stain. In general, 100ul of ready-to-use substrate solution is enough to cover one tissue slide.
Incubate for 5 mins at RT with the ready-to-use substrate solution. Rinse slides with deionized water.
Counterstain: Counterstain with hematoxylin. Rinse slides with deionized water.
Note:
Use TBS-T or PBS-T as wash buffer.
Storage:
Store at 2°C to 8°C, do not freeze.
Data reviews: side-by-side comparison
Manual Staining Procedure：
Primary antibody incubation: RT, 60min
Secondary antibody incubation: RT, 30min
DAB incubation：5min
Detection polymers in comparison:
MicroStackerTM Polymer Detection Kit, Universal
DAKO REAL EnVision Detection System
