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Home Product IMMUNOHISTOCHEMISTRY MICROSTACKER™ DETECTION SYSTEMS MicroStacker™ Polymer Detection Kit Universal

MicroStacker™ Polymer Detection Kit Universal

MicroStacker™ Polymer Detection System Universal is a biotin-free, polymeric horseradish peroxidase (HRP)- secondary antibody conjugate system for the detection of mouse and rabbit primary antibody on formalin-fixed, paraffin-embedded (FFPE) tissues in an immunohistochemistry (IHC) procedure. The innovative MicroStacker™ technology allows well-controlled layered stacking of antibodies and peroxidase enzymes on a micro-polymer scaffold. This technique results in a compact polymeric structure that easily penetrates to all cellular compartments, which provides superior sensitivity compared to other conventional HRP polymers with bulky dextran backbones. Plus, the system utilizes F(ab’) fragments of IgG secondary antibody instead of the whole IgG, which avoids the background caused by non-specific binding of whole IgG to endogenous Fc receptors.

Product Features

 

  • Known Applications

Immunohistochemistry (formalin-fixed paraffin-embedded tissues)

 

  • Scheme

 

  • 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

 

Experimental Results:

Breast (Her-2 / rabbit monoclonal antibody)

REAL EnVision Detection System

MicroStackerTM Polymer Detection, Universal

 

Ovary (CK5 / rabbit monoclonal antibody)

REAL EnVision Detection System

MicroStackerTM Polymer Detection, Universal

 

Specification

Code

Product Name

Components

Specification

SD3100

MicroStackerTM Polymer Detection Kit, Universal (3 Components)

MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer

100ml(1000T)

20*DAB Chromogen

5ml

DAB Substrate Buffer

100ml

MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer

30ml (300T)

20*DAB Chromogen

1.5ml

DAB Substrate Buffer

30ml

MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer

10ml (100T)

20*DAB Chromogen

1ml

DAB Substrate Buffer

10ml

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