小鼠凝血因子Ⅷ相关抗原(FⅧ-Ag)ELISA试剂盒厂家

检测标本：血清,血浆,尿液,胸腹水,脑脊液,细胞培养上清,组织匀浆等

检测方法：ELISA

检测类型：酶联免疫夹心法

产品的用途：仅供科研究课题使用

价格及详细资料：齐一生物销售：021－6034 8496；181214 53965；173021 04490；.齐一生物科技（上海）有限公司提供的ELISA试剂盒受到了广大科研单位的一致肯定和认同.大品牌保证,价格公道,倾力为国内外科研院校实验室提供最优质的产品.若有需要,我司将竭诚为您服务！本试剂盒用于测定血清,血浆及相关液体样本中含量.

【小鼠凝血因子Ⅷ相关抗原(FⅧ-Ag)ELISA试剂盒厂家】实验原理：

   本试剂盒应用双抗体夹心法测定标本中该产品水平.用纯化的本产品抗体包被微孔板,制成固相抗体,往包被单抗的微孔中依次加入本产品抗原,再与HRP标记的本产品抗体结合,形成抗体-抗原-酶标抗体复合物,经过彻底洗涤后加底物TMB显色.TMB在HRP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本产品呈正相关.用酶标仪在450nm波长下测定吸光度（OD值）,通过标准曲线计算样品中该产品浓度.

试剂盒组成：

齐一生物销售：021－6034 8496；181214 53965；173021 04490

【小鼠凝血因子Ⅷ相关抗原(FⅧ-Ag)ELISA试剂盒厂家】标本要求：

1．标本采集后尽早进行提取,提取按相关文献进行,提取后应尽快进行实验.若不能马上进行试验,可将标本放于-20℃保存,但应避免反复冻融

2．不能检测含NaN3的样品,因NaN3抑制辣根过氧化物酶的（HRP）活性.

操作步骤：

1.     标准品的稀释与加样：在酶标包被板上设标准品孔10孔,在第一、第二孔中分别加标准品100μl,然后在第一、第二孔中加标准品稀释液50μl,混匀；然后从第一孔、第二孔中各取100μl分别加到第三孔和第四孔,再在第三、第四孔分别加标准品稀释液50μl,混匀；然后在第三孔和第四孔中先各取50μl弃掉,再各取50μl分别加到第五、第六孔中,再在第五、第六孔中分别加标准品稀释液50ul,混匀；混匀后从第五、第六孔中各取50μl分别加到第七、第八孔中,再在第七、第八孔中分别加标准品稀释液50μl,混匀后从第七、第八孔中分别取50μl加到第九、第十孔中,再在第九第十孔分别加标准品稀释液50μl,混匀后从第九第十孔中各取50μl弃掉.（稀释后各孔加样量都为50μl,浓度分别为1200 ng/L,800 ng/L ,400 ng/L,200ng/L, 100 ng/L）. 【小鼠凝血因子Ⅷ相关抗原(FⅧ-Ag)ELISA试剂盒厂家】

2.     加样：分别设空白孔（空白对照孔不加样品及酶标试剂,其余各步操作相同）、待测样品孔.在酶标包被板上待测样品孔中先加样品稀释液40μl,然后再加待测样品10μl（样品最终稀释度为5倍）.加样将样品加于酶标板孔底部,尽量不触及孔壁,轻轻晃动混匀.

3.     温育：用封板膜封板后置37℃温育30分钟.

4.     配液：将30（48T的20倍）倍浓缩洗涤液用蒸馏水30（48T的20倍）倍稀释后备用.

5.     洗涤：小心揭掉封板膜,弃去液体,甩干,每孔加满洗涤液,静置30秒后弃去,如此重复5次,拍干.

6.     加酶：每孔加入酶标试剂50μl,空白孔除外.

7.     温育：操作同3.

8.     洗涤：操作同5.

9.     显色：每孔先加入显色剂A50μl,再加入显色剂B50μl,轻轻震荡混匀,37℃避光显色15分钟.

10. 终止：每孔加终止液50μl,终止反应（此时蓝色立转黄色）.

11. 测定：以空白空调零,450nm波长依序测量各孔的吸光度（OD值）. 测定应在加终止液后15分钟以内进行.

注意事项：

1． 试剂盒从冷藏环境中取出应在室温平衡15-30分钟后方可使用,酶标包被板开封后如未用完,板条应装入密封袋中保存.

2． 浓洗涤液可能会有结晶析出,稀释时可在水浴中加温助溶,洗涤时不影响结果. 【小鼠凝血因子Ⅷ相关抗原(FⅧ-Ag)ELISA试剂盒厂家】

3． 各步加样均应使用加样器,并经常校对其准确性,以避免试验误差.一次加样时间最好控制在5分钟内,如标本数量多,推荐使用排抢加样.

4． 请每次测定的同时做标准曲线,最好做复孔.如标本中待测物质含量过高（样本OD值大于标准品孔第一孔的OD值）,请先用样品稀释液稀释一定倍数（n倍）后再测定,计算时请最后乘以总稀释倍数（×n×5）.

5． 封板膜只限一次性使用,以避免交叉污染.

6． 底物请避光保存.

7． 严格按照说明书的操作进行,试验结果判定必须以酶标仪读数为准.

8． 所有样品,洗涤液和各种废弃物都应按传染物处理.

9． 本试剂不同批号组分不得混用.

10. 如与英文说明书有异,以英文说明书为准.

计算：

以标准物的浓度为横坐标,OD值为纵坐标,   

在坐标纸上绘出标准曲线,根据样品的OD     

值由标准曲线查出相应的浓度；再乘以稀释     

倍数；或用标准物的浓度与OD值计算出标     

准曲线的直线回归方程式,将样品的OD值     

代入方程式,计算出样品浓度,再乘以稀释

倍数,即为样品的实际浓度.

试剂盒性能：

1.样品线性回归与预期浓度相关系数R值为0.95以上.

2.批内与批见应分别小于9%和11%    

保存条件及有效期：

1.试剂盒保存；2-8℃.

2．有效期：6个月

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