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大鼠羟脯氨酸,大鼠羟脯氨酸试剂盒

简要描述:大鼠羟脯氨酸试剂盒 范围: 人,小大鼠,猪,兔,其它动物细胞因子;细胞凋亡,活性多肽,自身抗体 ,血栓与止血, 骨代谢,肝纤维化,肿瘤,激素内分泌,自身抗体科研ELISA检测试剂盒。

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  • 更新时间:2016-07-15
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详细介绍

   产品名称:大鼠羟脯氨酸试剂盒 
    产品规格:96人份/48人份
    :
    检测方法:酶联免疫法/酶免法(ELISA)
    96T指可以做94个标本,2个标准对照
    48T指可以做47个标本,1个标准对照
    96T-84个样本
    48T-42个样本
    该产品中英文操作步骤以人凝血酶激活纤溶抑制物试剂盒为例:
大鼠羟脯氨酸试剂盒  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*、第二孔中分别加标准品100μl,然后在*、第二孔中加标准品稀释液50μl,混匀;然 后从*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6μg/ml,4μg/ml ,2μg/ml,1μg/ml , 0.5μg/ml)。
    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zui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。
    Assay procedure
    1.Dilute and add sample to Standard: set 10 Standard wells on the ELISA plates coated, add Standard 100μl to the first and the second well, then add Standard dilution 50μl to the first and the second well, mix; take out 100μl form the first and the second well then add it to the third and the forth well separay. then add Standard dilution 50μl to the third and the forth well ,mix ; then take out 50μl from the third and the forth well discard, add 50μl to the fifth and the sixth well ,then add Standard dilution 50μl to the fifth and the sixth well, mix ; take out 50μl from the fifth and the sixth well and add to the seventh and the eighth well, then add Standard dilution 50μl to the seventh and the eighth well ,mix ; take out 50μl from the seventh and the eighth well and add to the ninth and the tenth well, add Standard dilution 50μl to the ninth and the tenth well, mix , take out 50μl from the ninth and the tenth well discard(add Sample 50μl to each well after Diluting ,(density: 6μg/ml,4μg/ml ,2μg/ml,1μg/ml , 0.5μg/ml)
    2.add sample:Set blank wells separay (blank comparison wells don’t add sample and HRP-Conjugate reagent, other each step operation is same). testing sample well. add Sample dilution 40μl to testing sample well, then add testing sample 10μl (sample final dilution is 5-fold), add sample to wells , don’t touch the well wall as far as possible, and Gently mix.
    3.Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37℃.
    4.Configurate liquid: 30-fold (or 20-fold)wash solution diluted 30-fold (or 20-fold) with distilled water and reserve.
    5.washing:Uncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer to every well, still for 30s then drain, repeat 5 times, dry by pat.
    6.add enzyme:Add HRP-Conjugate reagent 50μl to each well, except blank well.
    7.incubate:Operation with 3.
    8.washing:Operation with 5.
    9.color:Add Chromogen Solution A 50ul and Chromogen Solution B to each well, evade the light preservation for 15 min at 37℃
    10.Stop the reaction:Add Stop Solution50μl to each well, Stop the reaction(the blue color change to yellow color).
    11.assay:take blank well as zero , Read absorbance at 450nm after Adding Stop Solution and within 15min.
大鼠羟脯氨酸试剂盒  范围: 人,小大鼠,猪,兔,其它动物细胞因子;细胞凋亡,活性多肽,自身抗体 ,血栓与止血, 骨代谢,肝纤维化,肿瘤,激素内分泌,自身抗体科研ELISA检测试剂盒。

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