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A Monoclonal-Antibody-Based ELISA for the Detection of Human FADD (Fas-associated Death Domain)

A Monoclonal-Antibody-Based ELISA for the Detection of Human FADD (Fas-associated Death Domain)

Jianqiang Ye, Hongxia Shao, Dingyuan Ma, Aijian Qin, Bing Han, Faiz M M T Marikar, Wei Cheng, Xiaofeng Huang, Yudong Qiu, Qingang Hu, Zichun Hua

Biotechnol Appl Biochem. 2008 Jul;50(Pt 3):143-6.

PMID: 18167119

Abstract:

FADD (Fas-associated death domain) has been widely expressed in various tissues and its expression has been recently demonstrated to correlate with tumour progression and prognosis. Currently, measurement of FADD expression mainly depends on Western-blot or immunohistochemical approaches. To develop a conventional sandwich ELISA avenue for the detection of FADD protein to supplement Western blotting or immunohistochemistry, a series of mAbs (monoclonal antibodies) specific for FADD protein, designated 3A3, 3F9, 3G4, 4B9, 4G1, 7A8, 7B8 and 7F4, were produced by fusing mouse s/p20 myeloma cells with the spleen cells of a mouse immunized with the Escherichia coli-expressed recombinant His(6)-FADD protein. On the basis of the characterization of these mAbs, purified 3F9 was selected as the capture antibody and the biotin-conjugated 3A3 was selected as the detection antibody in sandwich ELISA. The limit of detection for the ELISA was 0.3 ng of purified His(6)-FADD (FADD tagged with hexahistidine), and it could detect both recombinant and native human FADD protein. Furthermore, the positive reaction of the ELISA could be blocked by rabbit anti-FADD sera. All of these results indicated that the ELISA developed in the present paper could be a promising tool for detection of FADD protein.

Chemicals Related in the Paper:

Catalog Number	Product Name	Price
IAR42412372	Anti-FADD antibody, Mouse monoclonal	Price
IAR42412374	Anti-FADD antibody produced in rabbit	Price
IAR42412385	Monoclonal Anti-FADD antibody produced in mouse	Price
IAR42412476	FADD, His tagged human	Price

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