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Home Product IMMUNOHISTOCHEMISTRY MICROSTACKER™ DETECTION SYSTEMS MicroStacker™ Rabbit-on-Rodent HRP-Polymer Detection Kit

MicroStacker™ Rabbit-on-Rodent HRP-Polymer Detection Kit

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. After mouse/rat serum cross-adsorption, MicroStacker™ Rabbit-on-Rodent HRP-polymer detection kit can significantly reduce non-specific staining caused by endogenous immunoglobulin. The kit is mainly used for rabbit monoclonal antibody/rabbit polyclonal antibody on mouse/rat tissues staining.

Product Features

  • 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.

 

Data reviews: side-by-side comparison

 

Staining Procedure

Primary antibody incubation: RT, 60min
Secondary antibody incubation: RT, 30min
DAB incubation:5min

 

Detection polymers in comparison:

MicroStackerTM Rabbit-on-Rodent HRP-Polymer Detection Kit

Rabbit-on-Rodent HRP-polymer (American Research Market)

 

Experimental Results:

Mouse liver (FAH rabbit monoclonal antibody)

 

                                                                                               Rabbit-on-Rodent HRP-polymer 

                                                                                                  MicroStackerTM Rabbit-on-Mouse

 

Specification

Code

Product Name

Components

Specification

RS3000

MicroStackerTM Rabbit-on-Rodent HRP-Polymer Detection Kit (3 Components)

MicroStackerTM Anti-Rabbit HRP-Polymer

100ml (1000T)

20*DAB Chromogen

5ml

DAB Substrate Buffer

100ml

MicroStackerTM Anti-Rabbit HRP-Polymer

30ml (300T)

20*DAB Chromogen

1.5ml

DAB Substrate Buffer

30ml

MicroStackerTM Anti-Rabbit HRP-Polymer

20ml (200T)

20*DAB Chromogen

1ml

DAB Substrate Buffer

20ml

RS3001

MicroStackerTM Rabbit-on-Rodent HRP-Polymer Detection Kit (4 Components)

Peroxidase Blocking Reagents

100ml

MicroStackerTM Anti-Rabbit HRP-Polymer

100ml (1000T)

20*DAB Chromogen

5ml

DAB Substrate Buffer

100ml

Peroxidase Blocking Reagents

20ml

MicroStackerTM Anti-Rabbit HRP-Polymer

20ml (200T)

20*DAB Chromogen

1ml

DAB Substrate Buffer

20ml

RS3002

MicroStackerTM Rabbit-on-Rodent HRP-Polymer Detection Kit (5 Components)

Peroxidase Blocking Reagents

100ml

MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer

100ml (1000T)

20*DAB Chromogen

5ml

DAB Substrate Buffer

100ml

Hematoxylin

100ml

Peroxidase Blocking Reagents

20ml

MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer

20ml (200T)

20*DAB Chromogen

1ml

DAB Substrate Buffer

20ml

Hematoxylin

20ml

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