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58 Lampiran 1
Nilai pH pemisahan whey Kode Sampel PE5 Hari ke-1 PE5 Hari ke-2 PE5 Hari ke-3 PE5 Hari ke-4 PE5 Hari ke-5 PE5 Hari ke-6 PE5 Hari ke-7 PE5 Hari ke-8 9B Hari ke-1 9B Hari ke-2 9B Hari ke-3 9B Hari ke-4 9B Hari ke-5 9B Hari ke-6 9B Hari ke-7 9B Hari ke-8 18B Hari ke-1 18B Hari ke-2 18B Hari ke-3 18B Hari ke-4 18B Hari ke-5 18B Hari ke-6 18B Hari ke-7 18B Hari ke-8 JR2 Hari ke-1 JR2 Hari ke-2 JR2 Hari ke-3 JR2 Hari ke-4 JR2 Hari ke-5 JR2 Hari ke-6 JR2 Hari ke-7 JR2 Hari ke-8 A11 Hari ke-1 A11 Hari ke-2 A11 Hari ke-3 A11 Hari ke-4 A11 Hari ke-5 A11 Hari ke-6 A11 Hari ke-7 A11 Hari ke-8
pH Awal 6.82 6.65 6.72 6.74 6.71 6.68 6.67 6.71 6.54 6.61 6.61 6.56 6.54 6.52 6.58 6.54 6.71 6.90 6.90 6.83 6,78 6.81 6.83 6.84 6.77 6.77 6.45 6.60 6.62 6.56 6.54 6.67 6.84 6.68 6.68 6.67 6.73 6.71 6.80 6.80
pH Pengasaman 4.74 4.73 4.66 4.66 4.65 4.67 4.79 4.73 4.67 4.64 4.50 4.68 4.70 4.72 4.68 4.68 4.71 4.56 4.64 6.62 4.62 4.64 4.66 4.70 4.64 4.60 4.76 4.90 4.76 4.72 4.67 4.72 4.67 4.71 4.64 4.78 4.40 4.73 4.67 4.77
pH Netralisasi 6.81 6.76 6.75 6.94 6.71 6.80 6.72 6.71 6.76 6.88 6.77 6.82 6.73 6.76 6.80 6.77 6.79 6.88 6.80 6.84 6.78 6.86 6.80 6.81 6.87 6.88 6.80 6.75 6.85 6.80 6.70 6.80 6.70 6.86 6.71 6.81 6.79 6.84 6.75 6.87
59 Lampiran 1
Lanjutan nilai pH pemisahan whey Kode Sampel A2 Hari ke-1 A2 Hari ke-2 A2 Hari ke-3 A2 Hari ke-4 A2 Hari ke-5 A2 Hari ke-6 A2 Hari ke-7 A2 Hari ke-8 2028 Hari ke-1 2028 Hari ke-2 2028 Hari ke-3 2028 Hari ke-4 2028 Hari ke-5 2028 Hari ke-6 2028 Hari ke-7 2028 Hari ke-8 2029 Hari ke-1 2029 Hari ke-2 2029 Hari ke-3 2029 Hari ke-4 2029 Hari ke-5 2029 Hari ke-6 2029 Hari ke-7 2029 Hari ke-8 2069 Hari ke-1 2069 Hari ke-2 2069 Hari ke-3 2069 Hari ke-4 2069 Hari ke-5 2069 Hari ke-6 2069 Hari ke-7 2069 Hari ke-8
pH Awal 6.65 6.67 6.60 6.69 6.60 6.66 6.63 6.60 6.82 6.65 6.72 6.74 6.71 6.68 6.67 6.71 6.54 6.61 6.61 6.56 6.54 6.52 6.58 6.54 6.71 6.90 6.90 6.83 6.78 6.81 6.83 6.84
pH Pengasaman 4.74 4.50 4.82 4.81 4.85 4.78 4.66 4.70 4.74 4.73 4.66 4.66 4.65 4.67 4.79 4.73 4.67 4.64 4.50 4.68 4.70 4.72 4.68 4.68 4.71 4.56 4.64 6.62 4.62 4.64 4.66 4.70
pH Netralisasi 6.84 6.79 6.88 6.89 6.95 6.81 6.86 6.91 6.81 6.76 6.75 6.94 6.71 6.80 6.72 6.71 6.76 6.88 6.77 6.82 6.73 6.76 6.80 6.77 6.79 6.88 6.80 6.84 6.78 6.86 6.80 6.81
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Lampiran 2 Rataan dan simpangan baku berat jenis (g/ml) kolostrum dan susu kambing Hari Pemerahan Hari Ke-1 Hari Ke-2 Hari Ke-3 Hari Ke-4 Hari Ke-5 Hari Ke-6 Hari Ke-7 Hari Ke-8
PE (3) 1.0493 1.0421 1.0317 1.0334 1.0310 1.0300 1.0310 1.0315
Bangsa Kambing (n) Jawarandu (3) 1.0392 1.0384 1.0345 1.0323 1.0350 1.0340 1.0330 1.0320
SAPE (3) 1.0465 1.0415 1.0376 1.0362 1.0368 1.0360 1.0360 1.0360
61 Lampiran 3
Tabel sidik ragam komposisi kolostrum dan susu kambing PE, Jawarandu dan SAPE a Kandungan bahan kering kolostrum dan susu Sumber Bangsa Kambing Hari Pemerahan Interaksi Galat Total
DB 2 7 14 48 71
JK 65.42 4764.71 390.77 682.93 5903.84
KT 32.71 680.67 27.91 14.22
F 2.30 47.84 1.96
P 0.1113 0.0001 0.0426
b Kandungan bahan kering tanpa lemak kolostrum dan susu Sumber Bangsa Kambing Hari Pemerahan Interaksi Galat Total
DB 2 7 14 48 71
JK 8.83 398.39 24.21 69.02 500.47
KT 4.41 56.91 1.72 1.43
F 3.07 39.58 1.20
P 0.0555 0.0001 0.3045
F 5.18 43.11 2.23
P 0.0092 0.0001 0.0203
c Kandungan lemak kolostrum dan susu Sumber Bangsa Kambing Hari Pemerahan Interaksi Galat Total
DB 2 7 14 48 71
JK 83.03 2418.03 249.73 384.64 3135.44
KT 41.51 345.43 17.83 8.01
d Kandungan protein kolostrum dan susu Sumber
DB
JK
KT
F
P
Bangsa Kambing Hari Pemerahan Interaksi Galat Total
2 7 14 48 71
535.28 744.83 649.19 162.49 2091.79
267.64 106.40 46.37 3.38
79.06 31.43 13.70
0.0001 0.0001 0.0001
F 79.06 31.43 13.70
P 0.0001 0.0001 0.0001
e Kandungan laktoferin kolostrum dan susu Sumber Bangsa Kambing Hari Pemerahan Interaksi Galat Total
DB 2 7 14 48 71
JK 535.28 744.83 649.19 162.49 2091.79
KT 267.64 106.40 46.37 3.38
62
Lampiran 4
Kurva volume efluen pada fraksi protein susu kambing PE
63
Lampiran 4 Lanjutan kurva volume efluen pada fraksi protein susu kambing PE
64
Lampiran 4 Lanjutan Kurva volume efluen pada fraksi protein susu kambing PE
65
Lampiran 5
Kurva volume efluen pada fraksi protein susu kambing Jawarandu
66
Lampiran 5
Lanjutan kurva volume efluen pada fraksi protein susu kambing Jawarandu
67
Lampiran 5
Lanjutan kurva volume efluen pada fraksi protein susu kambing Jawarandu
68
Lampiran 6
Kurva volume efluen pada fraksi protein susu kambing SAPE
69
Lampiran 6
Lanjutan kurva volume efluen pada fraksi protein susu kambing SAPE
70
Lampiran 6 Lanjutan kurva volume efluen pada fraksi protein susu kambing SAPE
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Lampiran 7 Berbagai peralatan untuk penelitian a Alat Sentrifugasi dingin Hettich Zentrifugen Mikro 200R
b
Alat Hi – Trap Q-SP Anion Exchange Chromatoghraphy
c
Alat Corning Sterile Syringe Filter 0,20 µm
d
Alat Elektroforesis Bio Rad Mini Protean®3 System
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Lampiran 8
Bahan-bahan untuk analisis laktoferin dengan SDS-PAGE
1. Akrilamid/Bis 30% Akrilamidat sebanyak 30 g dan N,N,-Bis-methylene-acrylamide sebanyak 0.8 g dilarutkan dalam 100 ml akuades, disaring dan disimpan pada suhu 4 oC. 2. Tris-HCl 0.5 M pH 8.8 Tris Base sebanyak 6.06 g dilarutkan dalam 40 ml akuades. Nilai pH diatur sehinggga 8.8 dengan HCl 1 N. Volume larutan ditepatkan hingga 100 ml dengan akuades dan simpan pada suhu 4 oC. 3. Tris-Glycine pH 8.3 Tris Base sebanyak 12 g dan 57.6 g glycine dilarutkan dalam 1 900 ml akuades. Diatur pH hingga 8.3 dengan HCl 1 N. Tepatkan hingga 2 000 ml dengan akuades dan simpan pada suhu 4 oC. 4. Sodium Dodesil Sulfat (SDS) 10% SDS sebanyak 10 g dilarutkan ke dalam 75 ml akuades, diaduk perlahan hingga homogeny. Tepatkan hingga 100 ml dengan akuades. 5. Ammonium Persulphate Ammonium Persulphate sebanyak 0.5 g dilarutkan ke dalam 4.5 ml akuades. Larutan ini harus selalu dibuat baru setiap pengujian. 6. Larutan Fiksasi 12% Trichloroacetic acid (TCA) 7. Larutan Pewarna (Staining) Coomassie Blue R-250 sebanyak 0.125 g ditambahkan ke dalam 1 000 ml larutan methanol:akuades:asam asetat (5:4:1). 8. Larutan Destaining Metanol Methanol:akuades:asam asetat (5:4:1) 9. Komposisi Reservoir Buffer pH 8.3 Nama Bahan Akuades TrisGlycine stock Sodium Dodesil Sulfat (SDS) 10%
Jumlah 8 900 ml 1 000 ml 100 ml
73
10. Komposisi Dissociation Buffer Nama Bahan Akuades Tris-HCl 0.5 M pH 8.8 Glycerine Sodium Dodesil Sulfat (SDS) 10% Mercapto ethanol Bromophenolblue 5% (b/v)
Jumlah 10.0 ml 5.0 ml 5.0 ml 10.0 ml 0.5 ml 0.5 ml
11. Komposisi Running Gel Nama Bahan Akuades Tris-HCl 3.0 M pH 8.9 Sodium Dodesil Sulfat (SDS) 10% Akrilamida/Bis 30% Ammonium persulfat 10% N'N'N'N Tetramethylethylene diamine (TEMED)
Gel 7,5% 24.4 ml 5.0 ml 0.4 ml 10 ml 0.4 ml 0.02 ml
Gel 10% 20.4 ml 5.0 ml 0.4 ml 14 ml 0.4 ml 0.02 ml
12. Stacking Gel 3% Nama Bahan Akuades Tris-HCl 0.5 M pH 7.0 Sodium Dodesil Sulfat (SDS) 10% Akrilamida/Bis 30% Ammonium persulfat 10% N'N'N'N Tetramethylethylene diamine (TEMED)
Jumlah 7.54 ml 1.25 ml 0.1 ml 1.0 ml 0.1 ml 0.005 ml
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Lampiran 9 Grafik normalitas bobot molekul primer protein standar bangsa kambing
a
b
Kambing bangsa PE
Kambing bangsa Jawarandu
75
c
Kambing bangsa SAPE
