齐一供应人组织相容性复合体Ⅰ类相关基因D（MICD）试剂盒

齐一供应人组织相容性复合体Ⅰ类相关基因D（MICD）试剂盒有效期：6个月
预期应用：ELISA法定量测定血清、血浆、细胞培养上清或其它相关生物液体中的含量.
说明
1．试剂盒保存：-20℃（较长时间不用时）；2-8℃（频繁使用时）.
2．浓洗涤液低温保存会有盐析出,稀释时可在水浴中加温助溶.
3．中、英文说明书可能会有不一致之处,请以英文说明书为准.
4．刚开启的酶联板孔中可能会含有少许水样物质,此为正常现象,不会对实验结果造成任何影响.
齐一供应人组织相容性复合体Ⅰ类相关基因D（MICD）试剂盒实验原理
用纯化的抗体包被微孔板,制成固相载体,往包被抗该指标抗体的微孔中依次加入标本或标准品、生物素化的抗该指标抗体、HRP标记的亲和素,经过彻底洗涤后用底物TMB显色.TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色.颜色的深浅和样品中的该指标呈正相关.用酶标仪在450nm波长下测定吸光度（OD值）,计算样品浓度.
试剂盒组成及试剂配制
1. 酶联板（Assay plate ）：一块（96孔）.
2. 标准品（Standard）：2瓶（冻干品）.
3. 样品稀释液（Sample Diluent）：1×20ml/瓶.
4. 生物素标记抗体稀释液（Biotin-antibody Diluent）：1×10ml/瓶.
5. 辣根过氧化物酶标记亲和素稀释液 （HRP-avidin Diluent）：1×10ml/瓶.
6. 生物素标记抗体（Biotin-antibody）：1×120μl/瓶（1：100）
7. 辣根过氧化物酶标记亲和素（HRP-avidin）：1×120μl/瓶（1：100）
8. 底物溶液（TMB Substrate）：1×10ml/瓶.
9. 浓洗涤液（Wash Buffer）：1×20ml/瓶,使用时每瓶用蒸馏水稀释25倍.
10. 终止液（Stop Solution）：1×10ml/瓶
10、标本的采集及保存
1. 血清：全血标本请于室温放置2小时或4℃隔夜后于1000 x g离心20分钟,取上清即可检测,或将标本放于-20℃或-80℃保存,但应避免反复冻融.
2. 血浆：可用EDTA或肝素作为抗凝剂,标本采集后30分钟内于2 - 8° C 1000 x g离心15分钟,或将标本放于-20℃或-80℃保存,但应避免反复冻融.
3. 细胞培养物上清或其它生物标本：1000 x g离心20分钟,取上清即可检测,或将标本放于-20℃或-80℃保存,但应避免反复冻融.
注：标本溶血会影响最后检测结果,因此溶血标本不宜进行此项检测.
标本的稀释原则：
首先通过文献检索的方式了解待测样本的大致含量,确定适当的稀释倍数.只有稀释至标准曲线的范围内,检测的结果才是准确的.稀释的过程中,应做好详细的记录.最后计算浓度时,稀释了“N”倍,标本的浓度应再乘以“N”.
标准品的稀释原则：2瓶,每瓶临用前以样品稀释液稀释至1ml,盖好后静置10分钟以上,然后反复颠倒/搓动以助溶解,其浓度为300 pg/ml,做系列倍比稀释后,分别稀释300 pg/ml,150 pg/ml,75 pg/ml,37.5 pg/ml,18.5 pg/ml,9 pg/ml,4.5 pg/ml,样品稀释液直接作为标准浓度0 pg/ml,临用前15分钟内配制.
如配制150 pg/ml标准品：取0.5ml（不要少于0.5ml）300 pg/ml的上述标准品加入含0.5ml样品稀释液的Eppendorf管中,混匀即可,其余浓度以此类推.
生物素标记抗体的稀释原则：
临用前以生物素标记抗体稀释液稀释,稀释前根据预先计算好的每次实验所需的总量配制（每孔100μl）,实际配制时应多配制0.1-0.2ml.如10μl生物素标记抗体加990μl生物素标记抗体稀释液的比例配制,轻轻混匀,在使用前一小时内配制.
辣根过氧化物酶标记亲和素的稀释原则：
临用前以辣根过氧化物酶标记亲和素稀释液稀释,稀释前根据预先计算好的每次实验所需的总量配制（每孔100μl）,实际配制时应多配制0.1-0.2ml.如10μl辣根过氧化物酶标记亲和素加990μl辣根过氧化物酶标记亲和素稀释液 的比例配制,轻轻混匀,在使用前一小时内配制.
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