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All Journal Jurnal Ilmu Ternak Veteriner Journal of the Indonesian Tropical Animal Agriculture
E.T. Setiatin
Faculty of Animal and Agricultural Sciences, Diponegoro University, Tembalang Campus, Semarang 50275, Central Java
Agriculture, Biological Sciences & Forestry Earth & Planetary Sciences Veterinary

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Extraction and isolation of Ovine Pregnancy-Associated Glycoprotein (ovPAG) from cotyledon placenta of Garut sheep Setiatin, E.T.; Sajuthi, D.; Purwantara, B.; Talib, C.
Indonesian Journal of Animal and Veterinary Sciences Vol 14, No 3 (2009)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (275.806 KB) | DOI: 10.14334/jitv.v14i3.342

Abstract

Pregnancy-associated glycoprotein (PAG) structurally related to aspartic protease, expressed in the outer epithelial cell layer (trophectoderm) of ungulate placenta. Ovine PAG (ovPAG) synthesized by mono- and binucleic trophoblast before complete implantation at Day 14-15. Of this, ovPAG could be used as a marker for early pregnancy. The objective of study was to extract and isolate PAG from placenta of Garut Sheep collected at term and to characterize their molecular weight. The procedures included extraction of protein at neutral pH (cotyledon was thawed, minced, added PBS, blended and centrifuged), acidic (H3PO41M, pH 4,5; centrifuged) and ammonium sulfate (40% and 80% (NH4)SO4, centrifuged) precipitation; gel filtration (Sephadex-G75), anion exchange chromatography (DEAE- cellulose). Cotyledon extract was subjected to Sephadex-G75 and DEAE cellulose, and their fractions were measured their absorbances. Absorbances of Sephadex-G75 and DEAE fractions at peak were assayed for protein concentration (Bichinconinic protein assay). Continuously, these fractions were subjected to monogel SDS-PAGE and stained by Commassie Brilliant Blue. It was four different molecular weights isolating from cotyledon of Garut Sheep, namely 68.8, 36.04, 32.39 and 12.18  kDa. However, after chromatography anion exchange (DEAE-cellulose), only  three bands exist, those were  71,67;  33,64 and 30,86 kDa. Key words: Garut sheep, Pregnancy-Associated Glycoprotein (PAG), Cotyledone, SDS-PAGE, DEAE-cellulose
THE EFFECT OF Anredera cordifolia (Ten.) Steenis SUPPLEMENTATION ON UTERINE INVOLUTION PROCESS EVALUATED BY OESTRUS POST PARTUM BEHAVIOR AND FERNING Purwasih, R.; Setiatin, E.T.; Samsudewa, D.
Journal of the Indonesian Tropical Animal Agriculture Vol 39, No 1 (2014): (March)
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.39.1.17-22

Abstract

The aim of this study was to determine the effect of Anrederacordifolia (Ten.) Steenissupplementation on uterine involution process in rabbit. The research design applied was completelyrandomized design with 4 treatments and 3 replications (T0 = without Anredera cordifoliasupplementation; TI = Anredera cordifolia supplemented two days before parturition; T2 = Anrederacordifolia supplemented two days after parturition; T3 = Anredera cordifolia supplemented two daysbefore until two days after parturition). The results showed that Anredera cordifolia (Ten.) Steenissupplementation could accelerate post partum oestrus in does that were characterized by post partumoestrus behavior, ferning of saliva and cervical mucus. The best treatment was T3, that was four daysAnredera cordifolia supplementation that was administered at 0.45 g/kg of body weight/day. Furtherresearch was needed to find an optimal dose of Anredera cordifolia (Ten.) Steenis on uterine involution.
ANTIBODY POLYCLONAL PRODUCTION ON RABBIT ANTI-OVINE PREGNANCY-ASSOCIATED GLYCOPROTEIN (Rabbit anti-ovPAG) Setiatin, E.T.; Sajuthi, D.; Purwantara, B.; Talib, C.; Fathul, F.; Adiati, U.; Praira, W.
Journal of the Indonesian Tropical Animal Agriculture Vol 36, No 3 (2011): (September)
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.36.3.159-165

Abstract

The aim of the study was to produce polyclonal antibody (rabbit anti-ovPAG) which could detectPAG in the urine of pregnant ewes. Twelve rabbits were immunized against ovPG DEAE-TrisHCl (DT),DEAE-NaCl 20mM (DN2), DEAE-NaCl 40mM (DN4), DEAE-NaCl 80mM (DN8), DEAE-NaCl160mM (DN16), DEAE-NaCl 320mM (DN32) and DEAE-NaCl 1M (DN1) and NaCl 0.9 % as aplacebo. The 0.5 ml of isolate (purified from ovine cotyledon) was emulsified in equal volume withcomplete and incomplete Freud’s adjuvant. The mixture of each isolate and adjuvant was injected atmutiple sites along the dorsal area of rabbits by subcutaneous route. Blood were collected from marginalear vein, starting before first injection (baseline) and every 14 days. Rabbit anti-ovPAG were measuredusing Modified ELISA Technique. By using Western Blot Technique, DN32 showed the best immuneresponse among others and also could differenciate ovPAG in the urine of pregnant ewes It could beconcluded that ovPAG DN32 is a specific source of rabbit anti-ovPAG production. Protein of ovPAG atmolecular weight 31 kDa is a pregnancy protein marker of garut sheep and could be developed as amajor protein for producing antibodi.
Co-Authors B. Purwantara C. Talib D. Sajuthi D. Samsudewa F. Fathul Rita Purwasih U. Adiati W. Praira