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Korte samenvatting Biochemie & Eiwittechnologie P. Cauwenberghs 2de jaar BMLT
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Korte samenvatting Biochemie & Eiwittechnologie P. Cauwenberghs 2de jaar BMLT
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1 Inleiding2 Biomoleculen
2.1 Water
2.2 Anorganische bestanddelen
2.3 Organische moleculen
2.4 Gluciden
- Polysaccariden
o Pectine
o Lignine
o Chitine
- Glycoproteinen en glycolipiden
o O of N gebonden
o Functies (SVTHR)
Stabiliseren EW
Vouwing EW vereenvoudigen
Targetting van EW
Celherkenning en receptorbinding
- Aanverwante moleculen
o Glycerol (propaan-1,2,3-triol)
o Ethyleenglycol (antivries)
o PEG (polyethyleenglycol = macrogol = laxeermiddel)
o Pyruvaat (pyrodruivenzuur)
o Lactaat (melkzuur)
o Glyceraldehyde-3-fosfaat
- Biologische rol (KERS RHW)
2.5 Lipiden
- Apolair
- Amfipatisch
2.6 Homeostase
3 pH en buffermengsels
3.1. PBS = phosphate buffered saline
3.2. Tris/HCl = tris hydroxymethylaminomethaan
3.3. EDTA = ethyleen diamine tetra acetaat
4. Biochemische technieken
4.1. Elektroforese
- Soorten dragers
o Cellulose/papier
o Celluloseacetaat
o Agar
o Agarose
o Polyacrylamide
o Capillair
- Kleuring
o Coomassie Briljant Blue R250 (EW)
o Ethidiumbromide (NZ)
4.1.1. Cellulose acetaat elektroforese
4.1.2. PAGE (SDS-PAGE)
o Acrylamide mbv cross-linker amoniumpersulfaat APS + TEMED tetramethylethyleendiamine
4.1.3. Agarose GE
, o NZ analyse tot 60 kB
o NZ analyse pulse field elektroforese > 2000 kb
o Iso-enzym patroon opsporen
4.1.4. Immunofixatie (AGE + immunoprecipitatie)
4.1.5. Iso-elektrische focussering (IEF)
o + 0,1M H2SO4 en - 1M NaOH
o Bepaling HbA1c (diabetes)
o Opsporen MS in cerebrospinaal vocht
o Opsporen EPO
4.1.6. 2D GE
4.1.7. Capillaire elektroforese (CE)
o Elektroforetische activiteit (gewoon) en elektro-osmotische flow EOF
o CDT (carbohydraat deficiënte transferrine) bepaling verhoging chr alcoholgebruik
o Serumpathologiën
4.2. Immunoblotting = westernblotting
o Nitrocellulosemembraan
4.3. Kolomchromatografie
4.3.1. Gelfiltratie
4.3.2. Ionwisselingschromatografie
4.3.3. Affiniteitschromatografie
o His-tag EW zuivering met IMAC = immobilized Metal Affinity Chromatography
4.4. Uit en inzouten van EW
4.5. Dialyse
4.6. Centrifugering
o Differentiële
o Zone
o Isopyknische centrifugering
4.7. Celfractionering
1) Isoleren en verkleinen weefsel
2) Openbreken van cellen en homogeniseren
3) Grove filtratie door gaas
4) Fractioneren van celcomponenten door centrifugeren (differentiële)
5) Verdere opzuivering en analyse
5. Proteïnen
Zie cursus
6. Enzymen en coënzymen
Zie cursus
7 Overzicht van het metabolisme
7.1 Anabolisme/katabolisme
7.2 Energiecycli id cel
7.2.1 ATP/ADP cyclus
- Tekening ATP
7.2.2 NAD+/NADH + H+ cyclus
- NADH is gereduceerde vorm van NAD+
- reductie en oxidatie
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