Product name

iVision^TM Poly-HRP Goat anti-Mouse, Rabbit IgG

Packing specification

Intended use

For Research Use only. iVision^TM Goat anti-Mouse, Rabbit IgG, peroxidase conjugated intended for the chromogenic detection of targeted antigens that have been reacted to a user-supplied primary antibody on human tissue. It is recommended that the reagents may not be substituted across kits or lot numbers.

Performance Characteristics

The detection systems have been found to be sensitive and show specific binding to the primary antibody with minimal to no non-specific binding. iVision^TM Detection systems have shown reproducible and consistent within a single run, between runs and between lots.  The products have been determined to be stable for the periods specified by real time testing.

Principle

Antigens in tissues and cells are detected in immunohistochemistry (IHC) by a two-stage process: the binding of a primary antibody and the subsequent detection of this binding by iVision^TM Detection system. Tissues are sectioned and mounted on a slide. Primary antibody produced in mouse is then applied to react with targeted antigens followed by the application of Poly-HRP Goat anti-Mouse, Rabbit IgG to react with primary antibodies, and finally, a substrate/chromogen solution, such as DAB or AEC, which is converted by peroxidase into insoluble deposit. The whole procedure generates a visible profile of locations of antigens in tissue sections or in types of cells.

Main components

Goat anti-Mouse, Rabbit IgG, Peroxidase Conjugated

Storage

Store at 2～8℃ for 18 months.

Sample requirements

FFPE tissues are usually cut into sections as thin as 3～5μm with a microtome. These sections are then mounted onto glass slides that are coated with a tissue adhesive.

Protocol

1. Sample preparation：Deparaffinize the slides in xylene Ⅰ, Ⅱ, Ⅲ for 5 minutes；Transfer the slides once through 100%, 100%, 95%, 75% alcohols for 2 minutes respectively. Rinse slides with deionized water for 30 seconds.

2. Blocking：Block endogenous peroxidase activity by incubating sections in 3% H₂O₂ solution at room temperature for 5 minutes to block endogenous peroxidase activity. Rinse the slides with deionized water for 30 seconds.

3. Antigen retrieval：Heat the EDTA Antigen retrieval buffer to 100℃. Then place the slides in the boiled buffer and continue to heat for 15～20 min. Naturally cool down for 30 minutes. Rinse the sample with wash buffer.

4. Primary antibody incubation: Drain the slides. Add primary antibody to tissue, incubate at room temperature for 30 minutes. (use antibody diluent or PBS as control). Wash the slides in PBST for 2 times, 5 minutes for each time. If the Primary antibody is concentrated, please dilute it to RTU(ready to use) according to the information on packing. 

5. Secondary antibody: Drain the slides. Add iVision^TM Poly-HRP Goat anti-Mouse, Rabbit IgG secondary antibody to tissue and incubate at room temperature for 20 minutes. Wash the slides in PBST for 2 times, 5 minutes for each time.

6. DAB：Drain the slides. Add DAB to the tissue and incubate at room temperature for 5 min. Rinse slides with deionized water.

7. Hematoxylin staining：Drain the slides. Add Hematoxylin to the tissue and incubate at room temperature for 5 minutes. Rinse slides with water. Use the acid solution for differentiation. Rinse slides with water.

8. Dehydrate：Dehydrate the slides in 75%, 95%, 100% alcohols for 2 minutes. Dry the slides. Cover stained tissue with a coverslip using mounting medium.

Expected Results

Proper use of this reagent will result in intense, clear staining at the antigen sites in both the specimen and positive control. Any deviation from these expected results should cause users to question the results and consult technical service or local distributor for assistance.

Precautions

1. Please read the instruction carefully and become familiar with all components of the kit prior to use, Strictly follow the instruction during operation.

2. DO NOT use the kit or any kit component after their.

3. Only trained professionals can use this kit. Please wear suitable lab coat and disposable gloves while handling the reagents.

4. Avoid contact of skin, eyes and mucous membranes with the chemicals.

5. DO NOT pipet by mouth.

6. Unused reagents, used kit and waste must be disposed according to local regulations.

Manufacturer

Company Name: Xiamen Talent Biomedical Technology Co.,Ltd

Address: No.3rd and 4th Floors, Building B10, No. 2068 Wengjiao Road West, BioMedical Park, Haicang District, Xiamen City, 36100 China

Tel: +86 592 6315755            

E-mail: tlsw@talentbiomedical.com                

Website: www.talentbiomedical.com

Symbols