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May I ask how much mouse liver tissue micro-sample is needed for BCA quantification?

Compliance
Asked by:AndyCell2587 2026-08-05 Views 216 4 answers
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All Answers4
K
KevinResearch 2026-09-21
裂解液影响要分开看。1% Triton X-100常在BCA可接受范围内,但一定要用同样的裂解液稀释BSA标准品,否则标准曲线会偏。SDS风险更大,尤其1% SDS已较高,容易产生沉淀或显色抑制,建议把样品至少稀释5-10倍使SDS降到0.1%以下,或选择说明书明确兼容高浓度SDS的BCA试剂盒。含DTT或还原剂时用兼容还原剂配方。
R
RexPlate601 2026-09-14
这周到货的话优先国产现货,我常用碧云天、雅酶、康为世纪的BCA试剂盒,本地代理或官方店问有没有库存;微量样买小包装即可,没必要为便宜买大包装。重点看产品说明里能否耐受你的裂解液浓度。如果急用,可要求供应商发顺丰或同城闪送,普通试剂常温通常没问题,但不要买临期或包装破损。
田
田艳4213 2026-09-24
小鼠肝匀浆一两百微升做BCA足够,别太担心。常规BCA每孔加20ul样品+200ul工作液,复孔2-3个也就几十微升。但肝组织蛋白浓度高,通常2-10 mg/ml,必须稀释10-50倍再测,否则超出线性。样品量特别少也可用半量法:10ul样品+100ul工作液。离心后取上清,定量前先稀释一批,避免反复冻融。
学
学生君 2026-08-11
肝组织匀浆后建议4℃ 12000g离心10-15min,取中间清液,避开上层油脂和底部沉淀。肝脏含血较多,匀浆液颜色深会抬高背景,离心不能完全去掉红色时可在标曲中加入相同裂解液并做样品空白。为提高低量样品的可靠性,可先做Bradford粗估浓度,再用BCA精确定量;Western上样前按BCA结果归一化。
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