Description
Introduction: Histatins are human histidine-rich and mostly cationic proteins identified originally by Oppenheim et al (1988) as antimicrobial and fungistatic peptides in human parotid and submandibular-sublingual gland secretions. Histatins possess antimicrobial activity against a broad spectrum of bacteria and fungi. Some histatins have been found to be binding proteins for tannins Histatin-5 has been shown to be a strong inhibitor of trypsin-like protease produced by Bacteroides gingivalis and also inhibits Clostripain. Gusman et al (2001) have reported that histatin-5 inhibits the activities of MMP-2 and MMP-9. In addition, Histatin is acftive against the human parasitic protozoa Leishmania.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to histatin-5. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for histatin-5 and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain histatin-5, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of histatin-5 in the samples is then determined by comparing the O.D. of the samples to the standard curve