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[Manual] MicroStacker™ Polymer Detection Kit Universal, 1 Step

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.

Recommended Protocol

Deparaffinization

Deparaffinize slides in xylene, then hydrate through graded alcohols to water.

Peroxidase Blocking

Apply Peroxidase Blocking Reagent for 5 minutes. Wash twice with wash buffer for 5 minutes each.

Antigen Retrieval

Perform antigen retrieval according to the primary antibody datasheet.

Primary Antibody

Apply approximately 100 μL, or sufficient volume to cover the tissue section. Incubate for 30 to 60 minutes at room temperature. Wash twice with wash buffer for 5 minutes each.

Detection Polymer

Apply approximately 100 μL of Anti-Mouse/Rabbit HRP-Polymer, or sufficient volume to cover the tissue section. Incubate for 30 minutes at room temperature. Wash twice with wash buffer for 5 minutes each.

DAB Development

Prepare the DAB working solution by mixing DAB Chromogen with DAB Substrate Buffer at a 1:20 ratio. Use approximately 100 μL per slide. Incubate for 5 minutes at room temperature, then rinse with deionized water.

Counterstaining

Counterstain with hematoxylin, then rinse with deionized water.

Specification

DAB Substrate Buffer

Code	Product Name	Components	Specification
SD3001	DAB Chromogen Kit	DAB Substrate (20x)	1000test/box
SD3001	DAB Chromogen Kit	Substrate Buffer	2000test/box
SD3100	MicroStacker^TM Polymer Detection Kit, Universal (3 Components)	MicroStacker^TM Anti-Mouse/Rabbit HRP-Polymer	100ml(1000T)
		20*DAB Chromogen	5ml
		DAB Substrate Buffer	100ml
		MicroStacker^TM Anti-Mouse/Rabbit HRP-Polymer	30ml (300T)
		20*DAB Chromogen	1.5ml
		DAB Substrate Buffer	30ml
		MicroStacker^TM Anti-Mouse/Rabbit HRP-Polymer	10ml (100T)
		20*DAB Chromogen	1ml
		DAB Substrate Buffer	10ml
SD3101	MicroStacker™ Polymer Detection Kit, Universal (4 components)	Peroxidase Block	1000test/box
		Anti-Mouse/Rabbit HRP-Polymer	1000test/box
		DAB Chromogen (20x)	2000test/box
		DAB Substrate Buffer	2000test/box
SD3102	MicroStacker™ Polymer Detection Kit, Universal (5 components)	Peroxidase Block	1000test/box
		Anti-Mouse/Rabbit HRP-Polymer	1000test/box
		DAB Chromogen (20x)	2000test/box
		DAB Substrate Buffer
		Hematoxylin

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Recommended Protocol
Specification

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