Isolation of lactating goat mammary plasma membranes
Sharma, A.; Ray, T.K.
Indian Journal of Experimental Biology 20(3): 257-259
1982
ISSN/ISBN: 0019-5189 PMID: 7118208 Document Number: 186229
Lactating goat mammary tissue obtained immediately post mortem was held in ice cold buffer (1 mM NaHCO3, 0.5 mM CaCl2, 0.09 M sucrose, pH 7.4), cut into approx. 1-cmsuperscript 3 pieces, dried and then minced with scissors until a paste was formed (approx. 2 h). The paste was stirred in buffer with a glass rod at low speed for 5 min and filtered through cheese cloth. The filtrate was then centrifuged repeatedly at low speed and the membrane fraction was purified by discontinuous sucrose density gradient centrifugation. The membrane fraction isolated by this technique was judged essentially pure ( more than 85%) on the basis of electron microscopic examination and activities of marker enzymes. Yield of plasma membranes using this system was 20-25 mg/100 g tissue.