Reagente de anticorpo PHH3 para imuno-histoquímica

【Product name】

IHC Antibody -- PHH3

【Packing specification】

【Intended use】

For Research use only. PHH3 Antibody reagent is intended for use to qualitatively identify PHH3 by microscopy in sections of FFPE tissue using immunohistochemical detection system.

【Principle】

Phosphohistone H3 (PHH3) is a core histone protein, which together with other histones, forms the major protein constituents of the chromatin in eukaryotic cells. Immunohistochemical studies showed anti-PHH3 specifically detected the core protein histone H3 only when phosphorylated at serine 10 or serine 28. 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. Desidratar : Desidratar as lâminas em álcool 75% , 95 % , 100%  por 2 minutos .  Seque as lâminas. Cubra o tecido manchado com uma lamínula usando meio de montagem.

【Localização positiva 】

1.  Localização positiva :  nuclear .

2.  Controle positivo :  tonsila .

【Precauções 】

1.  Leia atentamente as instruções e familiarize-se com todos os componentes do kit antes de usar. Siga rigorosamente as instruções durante a operação.

2.  NÃO use o kit ou qualquer componente do kit após a sua.

3.  Somente profissionais treinados podem usar este kit. Use jaleco adequado e luvas descartáveis ​​ao manusear os reagentes.

4.  Evite o contato da pele, olhos e membranas mucosas com os produtos químicos.

5.  NÃO pipete com a boca.

6.  Reagentes não utilizados, kits usados ​​e resíduos devem ser descartados de acordo com os regulamentos locais.

【Fabricante 】_

Nome da empresa: Xiamen Talent Biomedical Technology Co.,Ltd

Endereço: nº 3º e 4º andares, edifício B10, nº 2068 Wengjiao Road West, BioMedical Park, distrito de Haicang, cidade de Xiamen, 36100 China

Telefone: +86 592 6315755            

E-mail : tlsw@talentbiomedical.com                

Site : www.talentbiomedical.com

【 Símbolos 】