KESIMPULAN
1. Isolat ovPAG dapat diisolasi dari kotiledon plasenta yang dikoleksi sesaat setelah melahirkan
2. Respon imun terbaik berasal dari DN32
3. Rabbit anti-ovPAG DN32 dipergunakan sebagai sumber antibodi
4. Protein 31 kDa hanya terdapat pada urin domba bunting sedangkan protein sekitar 71 kDa sebagai protein umum
5. Pengenceran seri ovPAG DN32 dengan konsentrasi 0,344; 0,172; 0,0688; 0,0344 dan 0,0172 ng/ml dapat digunakan sebagai standar 6. Koefisien variasi intra-asai tinggi dan rendah adalah 9,97 % dan 5,87 % 7. Koefisien variasi inter-asai tinggi dan rendah adalah 16,95% dan
13,9 %.
8. Teknik ELISA Termodifikasi dapat membedakan ovPAG dalam urin domba bunting dan tidak bunting (55,56%).
SARAN
1. Perlu dilakukan purifikasi ulang ovPAG DN32 2. Perlu evaluasi dosis ovPAG yang diimunisasikan 3. Pemurnian rabbit anti-ovPAG DN32
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LAMPIRAN
Lampiran 2 Sodium Dedocyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-
PAGE)
RUNNING GEL, MONOGEL SDS-PAGE
No. Reagen 7.5% 10% 12% 14% 15% 1 Acrylamide 40% 2.73 mL 3.65 mL 4.38 mL 5.1 mL 5.47 mL 2 Bis- Acrylamide 2% 1.50 mL 2.01 mL 2.40 mL 2.80 mL 3.02 mL 3 SDS 10% 150.00 µL 150.00 µL 150.00 µL 150.00 µL 150.00 µL 4 Tris-HCl pH : 8.8 3.75 mL 3.75 mL 3.75 mL 3.75 mL 3.75 mL 5 Aquadest 6.78 mL 5.40 mL 4.24 mL 3.12 mL 2.53 mL 6 TEMED 7.50 µL 7.50 µL 7.50 µL 7.50 µL 7.50 µL 7 APS 10% 75.00 µL 75.00 µL 75.00 µL 75.00 µL 75.00 µL STACKING GEL 4% No. Reagen 4% 1 Acrylamide 40% 0.48 mL 2 Bis- Acrylamide 2% 0.26 mL 3 SDS 10% 50.00 µL 4 Tris-HCl pH : 6.8 1.26 mL 5 Aquadest 2.92 mL 6 TEMED 5.00 µL 7 APS 10% 25.00 µL RUNNING BUFFER 10x Trisma-base 30.3 g Glisin 144 g SDS 10 g Aquades hingga 1 L PH 8.3 SAMPLE BUFFER SDS10 % (b/v) 2 mL Tris-HCL PH 6,8 1.25 mL Gliserol 2.5 mL Brompenol biru 8.5% (b/v) 0.2 mL Aquades 3.55 mL B-mercaptoethanol 0.05 mL
BROAD RANGE MARKER, #161-0318
No. PROTEIN
BM (kDa)
1 Myosin 203.0
2 β-galactosidase 118.0 3 Bovine Serum Albumin 86.0 4 Ovalbumin 51.6 5 Carbonic Anhydrase 34.1 6 Soybean Trypsin Inhibitor 29.0 7 Lysozyme 19.2 8 Aprotivin 7.5
