Extract protein mMDH from pig heart

1. the preparation of pig heart
- Prepare potassium phosphate buffer pH = 7, 50mM
- 6,8g KH2PO4 + 50ml H20----> 1M KH2PO4
- 8,7 g K2HPO4 + 50 ml H2O -----> 1M K2HPO4
Dilute 20 times to have 50mM each of solution
Monitor pH by pH meter, Mix these solution, pour KH2PO4 solution into K2HPO4 and  until obtaining pH = 7.
Ratio 18(K2HPO4)/7(KH2PO4) = 2,57
- Pig hear stored at 4 degree C, is cut into small peaces, take 15g then but into 30 ml buffer containing MeSH(0,2% v/v = 60 ul). The use of MeSH is to ''break'' the bond S-S from protein, the tertiary structure and the quaternary structure of some proteins(including mMDH) can be disrupted from this process.
- Centrifugate the sample in 30 min with speed 11000.
- Collect the supertanent then filtered with 0.45um membrane.