VITELLOGENIN IDENTIFICATION AND PURIFICATION OF FOUR VARIANTS MONOMORPHIC ASIAN AROWANA : BANJAR (Scleropages macrocephalus), PAPUA (Scleropages jardinii), PINOH (Scleropages macrocephalus) AND SUPER RED (Scleropages legendrei), INITIAL STEP FOR SEX IDENTIFICATION PURPOSE

Arowana fish (Scleropages sp.) are monomorphic species, those animals that physically could not be  distinguished between male and female. Indonesia has five arowana species, those are Formosus (S. formosus), Super red (S. legendrei), Banjar and Pinoh (S. macrocephalus), Golden (S. aureus) and Papua (S. jardinii). Estradiol and Testosterone measurements was conducted to investigate whether the arowana male or female, but there was no significant relationship. Thus, the research was develoved through Vitellogenin (Vtg) detection. Vtg was confirmed as specific protein on female fish. Vitellogenin detection could be confirmed as female identification. In order to reach the validity of Vtg detection, the purified Vtg should be obtained from each species. The research was aimed to identified and purified Vitellogenin of four variants  Asian arowana : Banjar, Papua, Pinoh and Super red.

12 fishes from four variants of Arowana were induced with Estradiol 50 mg/kg bodyweight. The fishes were bleeding before and ten days after hormonal induction. The identification was performed using SDS-PAGE on estradiol induced fish plasma. The purification was performed using electro elution on 1,5 mm precut gel. The result of purification process was confirmed using SDS-PAGE on 0,75 mm gel. The concentration Each purified Vtg then quantified with A280 method using nanodrop.

The identification showed there were similiarity on Vtg of Banjar, Pinoh and Super red. The Vtg of these variants appeared in 180 kDa of molleculear weight (MW), while Vtg of Papua were consist of Vtg 1 and  Vtg 2 with 180 and 110 of MW resepectively. Purification only performed on three Variants : Banjar, Papuan and Super red. Quantification showed that the Vtg concentration of pure samples between 0,1 to 0,67 mg/mL. Pure vitellogenin of these variants were obtained and used for immunisation to obtain antibody and as antigen for Vtg detection immunoassay.