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A friend at a chicken farm is testing chicken serum and egg white protein. Is the BCA kit accurate?

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Asked by:刘辉宇 2026-06-27 Views 174 4 answers
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董
董文莉7052 2026-09-11
蛋清不要直接吸原液,粘稠会让枪头残留和气泡造成重复性很差。处理上我习惯先称重后加入含0.1% Triton X-100的PBS稀释10倍,涡旋后4℃ 12000g离心10min,取清液做检测;如果还拉丝,再过一次25G针头或短超声。做BCA时要设样品空白,因为稀释液中的去垢剂会抬高背景。每板都要带BSA标准曲线,不要用旧标曲。
B
Buffering7035 2026-09-13
禽类血清如果有溶血或脂血,BCA确实会受一点影响,但比Bradford温和。轻中度溶血会让结果略偏高,建议采血后尽快分离血清,再4℃ 10000g离心5-10min去掉脂层和细小沉淀。脂血较重的样本可以稀释2-5倍再测,标准曲线用同样稀释液。每批样本取2-3个做原倍、2倍、4倍平行稀释,如果线性不理想就加大离心或脱脂。蛋清特别粘,建议先加9倍PBS或生理盐水,用注射器来回吸打几十次或短暂超声,再离心取上清。
试
试剂老师 2026-09-16
BCA对还原剂、脂类的耐受有限,但动物血清里一般不含DTT或巯基乙醇,主要风险是脂血。如果只是测总蛋白,可以选增强型或兼容型BCA试剂盒,尤其说明可耐受1% Triton的国产品牌。品牌方面,我们实验室用过碧云天、康为世纪和索莱宝,常规动物样本差别不大;Thermo更稳但价格高、货期有时不稳。到货快优先找本地有现货的代理商,问清效期和批次。
A
AssayLine9227 2026-07-23
判断准不准不要只看说明书,建议拿3-5个代表性样本做加标回收和线性稀释。例如鸡血清分别原倍、2、4、8倍稀释,理论值应成比例;加一组BSA标准品到血清里看回收率,一般可接受范围在90-110%。如果脂血高,可以先高速离心脱脂或增大稀释倍数。蛋清也同样做5、10、20倍梯度,选落在线性中段的稀释倍数。试剂盒到货快可以选国产大包装,同城冷运比跨省更快。
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