Reactivo de anticuerpo HIF-1α para inmunohistoquímica

【Product name】

IHC Antibody --HIF-1α

【Packing specification】

【Intended use】

For Research use only. HIF-1α Antibody reagent is intended for use to qualitatively identify HIF-1α by microscopy in sections of FFPE tissue using immunohistochemical detection system.

【Principle】

Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response to hypoxia (1). The HIF1 complex consists of two subunits, HIF-1α and HIF-1β. HIF-1α plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Add the primary antibody to bind the antigen on tissue sections, and then use HRP labeled secondary antibody binding primary antibody to form the secondary antibody-primary antibody-antigen complex. When DAB chromogenic solution is added, HRP reacts with enzyme substrate to produce brown insoluble reaction product, which indirectly indicating the existence of antigen. 

【Main components】

Immunoglobulin, antibody diluent

【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: cytoplasm/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.  NO use el kit ni ningún componente del kit después de su.

3.  Solo los profesionales capacitados pueden usar este kit. Utilice una bata de laboratorio adecuada y guantes desechables mientras manipula los reactivos.

4.  Evite el contacto de la piel, los ojos y las membranas mucosas con los productos químicos.

5.  NO pipetear con la boca.

6.  Los reactivos no utilizados, el kit usado y los residuos deben eliminarse de acuerdo con las normativas locales.

【Fabricante 】_

Nombre de la empresa: Xiamen Talent Biomedical Technology Co., Ltd

Dirección: Pisos n.º 3 y 4, edificio B10, n.º 2068 Wengjiao Road West, parque biomédico, distrito de Haicang, ciudad de Xiamen, 36100 China

Teléfono: +86 592 6315755            

Correo electrónico : tlsw@talentbiomedical.com                

Sitio web : www.talentbiomedical.com

【 Símbolos 】