en 24285995 Background: Ferritin is an iron storage protein, which plays a key role in iron metabolism. Measurement of ferritin level in serum is one of the most useful indicators of iron status and also a sensitive measurement of iron deficiency. Monoclonal antibodies may be useful as a tool in various aspects of ferritin investigations. In this paper, the production of a murine monoclonal antibody (mAb) against human ferritin was reported. Methods: Balb/c mice were immunized with purified human ferritin and splenocytes of hyper immunized mice were fused with Sp2/0 myeloma cells. After four times of cloning by limiting dilution, a positive hybridoma (clone: 2F9-C9) was selected by ELISA using human ferritin. Anti-ferritin mAb was purified from culture supernatants by affinity chromatography. Results: Determination of the antibody affinity for ferritin by ELISA revealed a relatively high affinity (2.34×109 M-1) and the isotype was determined to be IgG2a. The anti-ferritin mAb 2F9-C9 reacted with 79.4% of Hela cells in flow cytometry. The antibody detected a band of 20 kDa in K562 cells, murine and human liver lysates, purified ferritin in Western blot and also ferritin in human serum. Conclusion: This mAb can specifically recognize ferritin and may serve as a component of ferritin diagnostic kit if other requirements of the kit are met. ELISA, Ferritin, Flow cytometry, Monoclonal antibody, Western blotting 212 219 https://www.ajmb.org/En/Article.aspx?id=127 https://www.ajmb.org/PDF/En/FullText/127.pdf Ali AhmadBayatMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran12OmidYeganehMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran600RoyaGhodsMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran8Amir-HassanZarnaniNanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran7RezaBahjati ArdekaniReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran604Ahmad RezaMahmoudiMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran33JafarMahmoudianMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran114FarzanehHaghighat-NoutashMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran609MahmoodJeddi-TehraniMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran15