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Fig. 7 | Molecular Medicine

Fig. 7

From: Purification and Characterization of Macrophage Migration Inhibitory Factor as a Secretory Protein from Rat Epididymis: Evidences for Alternative Release and Transfer to Spermatozoa

Fig. 7

Detection of MIF in the ODF of spermatozoa. (A) SDS-PAGE with subsequent Coomassie brillant blue staining of the purified outer dense fiber (ODF) proteins from epididymal spermatozoa. Characteristically are the prominent bands at around 80–85 kDa, a complex around 60–70 kDa, 47 kDa, and 23–32 kDa. A minor band was seen at around 12 kDa. (B) Western blots analysis of total sperm from caput and cauda epididymis, and isolated outer dense fiber (ODF) proteins obtained from epididymal spermatozoa show a strong immunoreactive band at 12 kDa when probed with the anti-MIF antibody. The supernatant yield from the second gradient centrifugation step, which is used to separate the ODF from other tail proteins, contains markedly less migration inhibitory factor (MIF). Recombinant MIF (rec MIF; 0.5 µg) was used as a positive control.

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Molecular Medicine

ISSN: 1528-3658