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Ultrasensitive Polymer-based LSAB IHC Kit (Rabbit) (100 Tests)

 Description: 

Immunohistochemistry (IHC) is a method of detecting the presence of specific proteins in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues. IHC is widely used in the diagnosis of abnormal cells and basic research to understand the distribution and localization of biomarkers and differentially expressed proteins in different parts of a biological tissue. 

This product is based on a novel LSAB (labeled streptavidin-biotin) method. In this new method a single primary antibody is subsequently associated with multiple biotin molecules. Poly-HRP Streptavidin provides multiple active biomolecules on each polymer chain which increase the biotin binding capacity and amplify the peroxidase enzyme signal

Contents: 

1. RTU normal goat serum: 10ml. 

2. RTU peroxidase blocking solution: 10ml 

3. RTU biotinylated anti-rabbit secondary antibody: 10ml 

4. RTU streptavidin-HRP polymer: 10ml 

Note: RTU=Ready-to-use 

Storage 

Store at 2-8°C. 


 Protocol: 

1. Preparation of Slides 

A. Cell Lines 

 Grow cultured cells on sterile glass cover slips or slides overnight at 37 º C 

 Wash briefly with PBS 

 Fix as desired. Possible procedures include: 

a. 20 minutes with 10% formalin in PBS (keep wet) 

b. 10 minutes with ice cold methanol, allow to air dry 

c. 10 minutes with ice cold acetone, allow to air dry 

 Wash in PBS 

B. Frozen Sections 

 Snap frozen fresh tissues in liquid nitrogen or isopentane pre-cooled in liquid nitrogen, embedded in OCT compound in cryomolds. Store the frozen tissue block at -80°C until ready for sectioning. 

 Transfer the frozen tissue block to a cryotome cryostat (e.g. -20°C) prior to sectioning and allow the temperature of the frozen tissue block to equilibrate to the temperature of the cryotome cryostat. 

 Section the frozen tissue block into a desired thickness (typically 5-10 μm) using the cryotome. 

 Place the tissue sections onto glass slides suitable for immunohistochemistry (e.g. Superfrost). 

 Sections can be stored in a sealed slide box at -80°C for later use. 



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