IHC-Tek Polymer-HRP Conjugated Goat Anti-Rabbit and Mouse IgG (H+L) Ready-to-Use

Product Description

Product Description / Introduction

This product utilize a dextran backbone to which multiple HRP enzyme and antibodies molecules are conjugated. This polymer creates a highly sensitive, readily available and one-step non-biotin detection system for immunohistochemical and immunocytochemical staining. This system avoids the use of streptavidin and biotin, and therefore eliminates non-specific staining as a result of endogenous biotin. It has a wide adaptability to sample processing, reducing the variability caused by different tissue processing methods. The reagent is ready-to-use, which does not need to be diluted to obtain the best staining ratio, reducing experimental operation errors.

Storage and Shipping Conditions

Wet ice packs for transportation, store at 2-8℃ for 12 months.

Assay Protocol / Procedures

This protocol is for manual staining reference only. For other instruments, refer to the instructions for each instrument.

Procedures	Reagent	Protocol
Dewax	Alcohol and Xylene	Standard de-wax steps
Antigen Retrieval	Antigen Retrieval (IW-1100-1L IHC-Tek Epitope Retrieval Solution)	Antigen Retrieval Heat at 95-100℃ for 30 min and cool for 20 min (IW-1102 IHC-Tek Steamer Set)
Blocking of the endogenous peroxidases	3% Hydrogen peroxide (IW-1300)	Incubate at room temperature for 10 min.
Serum Blocking	IHC-Tek Antibody Diluent	Not needed if primary antibody diluted with IW-1000 IHC-Tek Antibody Diluent
Primary antibody	Commercial Primary antibody	Working solution was incubated at 37℃ for 30-60 min and washed three times with PBS.
Secondary antibody	IHC-Tek Polymer-HRP conjugated antibody	incubated at RT for 20 min and washed three times with PBS.
DAB staining	DAB Substrate Kit	Add the prepared DAB working solution and develop color for 5-10 min, wash three times with water.
Hematoxylin staining	Mayer Hematoxylin Solution (IW-1400)	Incubate at room temperature for 1 min, Rinse 10 minutes with tap water.
Dehydration and Mounting	Mounting Medium	70% alcohol 5min - 95% alcohol 5min - absolute ethanol 5min x 2 - xylene 5min x 2，The section were removed from xylene and mounting.

Note

1. This product is a ready-to-use reagent without dilution.

2. If the background is high, the primary antibody can be diluted by gradient to obtain better results.,

3. Wear protective equipment. for your safety.

For Research Use Only!