Amphoteric Keratin was made by 3-chloro-2-hydroxypropyl trimethyl ammonium chloride and sodium chloroacetate.And study the Flocculation Effect on the Leather Dyeing Fatliquoring Wastewater in this paper.Laboratory work has been carried out to investigate the influences of amphoteric keratin dosage, pH value and flocculation time on the removal rate of CODcr and total Cr efficiency. Results showed that:when the flocculent dosage was 0.5g稬-1,pH 4.0 and flocculation time was 4h, removal of CODcr and total Cr from 18840mg稬-1,19.0225mg稬-1 to 3600mg稬-1 and 1.277mg稬-1,as well as the decolorization effect is obvious. The flocs were used to fill the chrome tanned leather after proper treatment.The results showed that the thickening rate was 7.32%,the tensile strengthen was strenghten as well as the elongation at break had good effect. what’s more ,it plays a good dispersion on the lether fibers and the reusing flocs are practial and effective.