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IMMUNOHISTOCHEMISTRY
MICROSTACKER™ DETECTION SYSTEMS
MicroStacker™ Polymer Detection Kit Enhanced 
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ZhengZhou (HQ), Shenzhen (R&D Center), Suzhou (R&D Center), China
MicroStacker™ Polymer Detection Kit Enhanced
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. Compared to the MicroStacker™ Polymer Detection System, Universal, the Enhanced kit comes with the HRP polymer at a higher degree of polymerization, thus higher sensitivity.
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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
Part 1:
- Manual Staining Procedure：
Primary antibody incubation: RT, 60min
Secondary antibody incubation: RT, 30min
DAB incubation：5min
- Detection polymers in comparison:
DAKO REAL EnVision Detection System
MicroStackerTM Polymer Detection Kit, Universal (MicroStacker, Universal)
MicroStackerTM Polymer Detection Kit, Enhanced (MicroStacker, Enhanced)
- Experimental Results：
Breast (Her-2 / rabbit monoclonal antibody)
REAL EnVision Detection System
MicroStacker, Universal
MicroStacker, Enhanced
Part 2:
- Manual Staining Procedure：
The staining protocol of “MicroStacker, Enhanced” and “REAL EnVision Detection System” are the same as part 1.
Protocol for “EnVision Flex + Mouse Linker”:
Primary antibody incubation: RT, 30min
Mouse Linker: RT, 20min
Secondary antibody incubation: RT, 20min
DAB incubation: 5min
- Detection Systems in Comparison:
MicroStacker, Enhanced
REAL EnVision Detection System
EnVision Flex + Mouse (Linker)
- Experimental Results:
Esophagus (D2-40 / mouse monoclonal antibody)
REAL EnVision
EnVision Flex + Mouse Linker
MicroStacker, Enhanced
Specification
|
MicroStackerTM Polymer Detection Kit Enhanced ,1-step
|
Code |
Product Name |
Components |
Specification |
|
SD3203 |
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 |
20ml (200T) |
|||
|
20*DAB Chromogen |
1ml |
|||
|
DAB Substrate Buffer |
20ml |
|||
|
SD3204 |
4 Components |
Peroxidase Blocking Reagents |
100ml |
|
|
MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer |
100ml |
|||
|
20*DAB Chromogen |
5ml |
|||
|
DAB Substrate Buffer |
100ml |
|||
|
Peroxidase Blocking Reagents |
20ml |
|||
|
MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer |
20ml |
|||
|
20*DAB Chromogen |
1ml |
|||
|
DAB Substrate Buffer |
20ml |
|||
|
SD3205 |
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 |
|
MicroStackerTM Polymer Detection Kit Enhanced ,2-step
|
SD3206 |
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 |
20ml (200T) |
|||
|
20*DAB Chromogen |
1ml |
|||
|
DAB Substrate Buffer |
20ml |
|||
|
SD3207 |
4 Components |
Peroxidase Blocking Reagents |
100ml |
|
|
MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer |
100ml |
|||
|
20*DAB Chromogen |
5ml |
|||
|
DAB Substrate Buffer |
100ml |
|||
|
Peroxidase Blocking Reagents |
20ml |
|||
|
MicroStackerTM Anti-Mouse/Rabbit HRP-Polymer |
20ml |
|||
|
20*DAB Chromogen |
1ml |
|||
|
DAB Substrate Buffer |
20ml |
|||
|
SD3208 |
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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