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ZhengZhou (HQ), Shenzhen (R&D Center), Suzhou (R&D Center), China
MicroStacker™ Polymer Detection System 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. 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. MicroStacker™ goat anti-rabbit HRP-polymer detection kit is mainly used for staining rabbit monoclonal antibody/rabbit polyclonal antibody on mouse/rat tissues and detection of rabbit antibodies.
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.
Primary antibody incubation: RT, 60min
Secondary antibody incubation: RT, 30min
DAB incubation: 5min
Data reviews: side-by-side comparison
Detection polymers in comparison:
MicroStackerTM Goat Anti-Rabbit HRP-Polymer Detection Kit
Rabbit-on-Rodent HRP-polymer (American Research Market)
Experimental Results:
Mouse kidney (EGFR)
Rabbit-on-Rodent
CNT
Code |
|
Product Name |
Components |
Specification |
RS1000 |
MicroStacker™ Goat Anti-Rabbit/Mouse Polymer Detection Kit
|
3 Components |
Anti-Rabbit/Mouse HRP-Polymer |
100ml (1000T) |
20*DAB Chromogen |
5ml |
|||
DAB Substrate Buffer |
100ml |
|||
Anti-Rabbit/Mouse HRP-Polymer |
30ml (300T) |
|||
20*DAB Chromogen |
1.5ml |
|||
DAB Substrate Buffer |
30ml |
|||
Anti-Rabbit/Mouse HRP-Polymer |
20ml (200T) |
|||
20*DAB Chromogen |
1ml |
|||
DAB Substrate Buffer |
20ml |
|||
RS1001 |
4 Components |
Peroxidase Blocking Reagents |
100ml |
|
Anti-Rabbit/Mouse HRP-Polymer |
100ml |
|||
20*DAB Chromogen |
5ml |
|||
DAB Substrate Buffer |
100ml |
|||
Peroxidase Blocking Reagents |
20ml |
|||
Anti-Rabbit/Mouse HRP-Polymer |
20ml |
|||
20*DAB Chromogen |
1ml |
|||
DAB Substrate Buffer |
20ml |
|||
RS1002 |
5 Components |
Peroxidase Blocking Reagents |
100ml |
|
Anti-Rabbit/Mouse HRP-Polymer |
100ml (1000T) |
|||
20*DAB Chromogen |
5ml |
|||
DAB Substrate Buffer |
100ml |
|||
Hematoxylin |
100ml |
|||
Peroxidase Blocking Reagents |
20ml |
|||
Anti-Rabbit/Mouse HRP-Polymer |
20ml (200T) |
|||
20*DAB Chromogen |
1ml |
|||
DAB Substrate Buffer |
20ml |
|||
Hematoxylin |
20ml |