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hshever

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[求助] 求翻译一段文献句子，关于蛋白质纯化的

Purification of gp96 and CRT.
A 40-ml Meth A cell pellet (or equivalent wet weight of mouse liver) was homogenized in 160 ml of hypotonic buffer (30 mM NaHCO3, 1 mM PMSF, pH 7.1)
and a 100,000g supernatant was obtained. Solid ammonium sulfate was added to bring the solution to 50% saturation. This was centrifuged at 14,000 rpm for 30 min. The precipitate was discarded and the supernatant was subjected to subsequent fractionation at 80% ammonium sulphate. After centrifugation at 14,000 rpm for 30 min the precipitate was solubilized in PBS containing 2 mM Ca2
and 2 mM Mg2. This was applied to a ConA–Sepharose column (Pharmacia). The Con A–bound proteins wereused for purification of gp96 as previously described (2). The buffer of Con A unbound material was changed to 25 mM Na citrate
buffer, pH 5.3, by PD-10 column (Sephadex G-25; Pharmacia Biotech.). It was then applied to the CM-Sephadex C-50 column. The buffer of unbound of CM-Sephadex was then changed to 19 mM Na-phosphate buffer, pH 6.1 by PD-10 columns. It was then applied to the DEAE-sephacel column and eluted with a linear
gradient of 0.15–0.5 M NaCl in 20 mM Na-phosphate buffer. The CRT-containing fractions were identified by immunoblots。

CRT是钙网蛋白特别是划线的句子，不是很明白

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