Reactivo de anticuerpo PCNA para inmunohistoquímica

【Nombre del producto 】

Anticuerpo IHC --  PCNA

【 Especificación de embalaje 】

【Uso previsto 】

Solo para uso de investigación. El reactivo PCNA Antibody está diseñado para identificar cualitativamente PCNA mediante microscopía en secciones de tejido FFPE utilizando un sistema de detección inmunohistoquímico.

【P rincipio 】

El PCNA (antígeno nuclear de células proliferantes) juega un papel central en la coordinación de la asociación de factores de replicación durante la replicación del ADN, así como durante el reconocimiento y la reparación del daño en el ADN. El anticuerpo PCNA detecta células en proliferación tanto en células normales como tumorales. Se ha demostrado que el índice de marcaje anti-PCNA está asociado con el pronóstico del tumor. Agregue el anticuerpo primario para unir el antígeno en las secciones de tejido y luego use el anticuerpo primario de unión del anticuerpo secundario marcado con HRP para formar el complejo anticuerpo secundario-anticuerpo primario-antígeno. Cuando se agrega la solución cromogénica DAB, la HRP reacciona con el sustrato enzimático para producir un producto de reacción insoluble de color marrón, lo que indica indirectamente la existencia del antígeno. 

【Componentes principales 】

Inmunoglobulina , diluyente de anticuerpos

【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 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.

【Positive localization】

1. Positive localization: nuclear.

2. Positive control: breast cancer.

【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

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