Question Detail
After preparing the LPS solution, store it at -20°C. Will repeated freeze-thaw cycles affect the effect when administered to mice?
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All Answers4
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WendyReagent4282
2026-09-20
LPS低温本身不容易化学降解,但反复冻融会改变其胶束结构,增加管壁吸附,造成实际注射量下降或局部浓度异常。如果实验死亡率波动大,可以做个简单控制:同批小鼠分别用新鲜配置和冻融3次的母液注射,观察体温和存活率差异。若不想额外做动物,也可用鲎试剂检测内毒素活性,但动物反应更直接。重点还是规范分装,不要把整瓶母液反复冻融,分装后每管只解冻一次。
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Tom7194
2026-09-14
冻存母液取出后放4℃或室温缓慢解冻,不要用热水或微波。完全融化后涡旋5-10秒,再短暂离心管壁,保证均一。如果发现溶液浑浊或有絮状,可能已聚集,不建议继续用于关键实验。腹腔注射前将所需母液用无菌无内毒素PBS稀释到工作浓度,稀释后2小时内用完。小鼠脓毒症模型剂量和死亡率关系密切,建议每次都从同一批分装管取用,保证实际注射剂量一致。
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DavidElisa4335
2026-09-23
保存最稳妥的做法:用无内毒素PBS或注射用水配母液,浓度不要过低,1mg/mL常用。分装前先涡旋充分,动作快一些,分装体积按每次实验所需量,如25μL/管或100μL/管,写上日期和浓度。短期几个月-20℃可以,长期或条件允许放-80℃更稳。避免使用反复开关造成温度波动的冰箱,不要把母液在4℃放置超过1周。工作液最好现配现用,注射前再从母液稀释到目标浓度。
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WangPCR
2026-09-13
LPS配成母液后反复冻融确实可能导致聚集和管壁吸附,尤其是低浓度时。死亡率波动大不一定全是LPS降解,更多是冻融后形成不同粒径的聚集体,注射后生物利用度不一致。我们实验室以前图省事冻融3次,结果同批小鼠反应差别很明显。建议配成1mg/mL母液后分装成单次用量,如50-100μL/管,-20或-80冻存,一次一管,不复冻。分装管用低吸附离心管,避免LPS贴壁损失。
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