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Peritoneal membrane isolation

VB Vikrant K. Bhosle
TM Tapas Mukherjee
YH Yi-Wei Huang
SP Sajedabanu Patel
BP Bo Wen (Frank) Pang
GL Guang-Ying Liu
MG Michael Glogauer
JW Jane Y. Wu
DP Dana J. Philpott
SG Sergio Grinstein
LR Lisa A. Robinson
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Serum samples were collected from C57BL/6J mice (ten males and ten females) as described before and animals were euthanized immediately after the blood collection. Under aseptic conditions, the abdominal cavity was opened by removing the skin and underlying muscle but keeping the peritoneal membrane intact. The peritoneal membrane was separated from underlying adipose tissue and organs and collected in cold PBS. The membrane was rinsed with PBS once and resuspended in RIPA buffer. Samples were homogenized with a tissue homogenizer and stored at −20 °C overnight. Two freeze-thaw cycles were performed to break cell membranes and homogenates were centrifuged at 5000 × g for 5 min at 4 °C. Supernatants were stored at −80 °C.

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