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Can the cell lysis buffer prepared with SDS still be used after being stored at 4°C for one week? Measuring chicken peripheral blood lymphocytes.

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Asked by:蔡燕7163 2026-08-10 Views 204 4 answers
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All Answers4
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Nina2956 2026-09-22
浑浊不一定是SDS析出,也可能是细菌或真菌污染,尤其裂解液含有一些营养成分。4度放一周有风险。我一般把配好的裂红液分装成10mL离心管,负20度避光保存,现取现用,避免反复冻融。如果开盖闻到酸味或氨味,或回温后仍浑浊,直接弃掉。做流式样本不要省这点试剂,标记抗体比裂解液贵很多,没必要为省50mL裂解液冒样本损坏的风险。
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小小同学7058 2026-09-09
4度放一周出现浑浊,很大可能是SDS低温下析出,并不是一定变质。先把管放25到37度水浴或室温回温,隔几分钟轻轻颠倒,如果浑浊消失、恢复澄清,说明是SDS析出,通常还能用。但裂解红细胞液里如果还含氯化铵或碳酸氢盐,放久了pH可能漂移,也可能长菌。回温后观察有无絮状物或异味,最好取少量试裂解正常红细胞,不要直接上正式样本。
M
Media 2026-09-20
可以用少量鸡外周血或红细胞悬液做对照:取少量裂解液和抗凝血混匀,看是否在1到2分钟内透亮,离心看沉淀颜色,再上流式看FSC/SSC碎片和淋巴细胞群比例。与新鲜配制的裂解液平行比较,如果裂解时间、细胞回收率、后续标记阳性率没有明显差异,可以继续用;如果裂解延迟或碎片增多,就弃掉换新。
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WendyPlate 2026-07-31
含SDS的工作液不建议4度放一周再用于正式实验。SDS本身在这个时间尺度上一般稳定,但配方里其他成分如氯化铵、EDTA、缓冲盐可能出现沉淀或pH变化,影响裂红效率。理想做法是配成10% SDS母液室温避光保存,其他成分配成2×母液4度保存,用前混合。如果已经放了一周,我建议重配,50mL成本不高,但流式样本和抗体更贵,不值得冒险。
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