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现在位置首页>实验试剂制品>生化试剂>ELISA试剂盒>人血纤肽试剂盒
人血纤肽试剂盒
人血纤肽试剂盒
  • 人血纤肽试剂盒

人血纤肽试剂盒

产品报价:询价

更新时间:2013/10/31 17:27:43

地:美国

牌:R&D

号: 96T

厂商性质: 生产型,

公司名称: 上海劲马实验设备有限公司

产品关键词:

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          人血纤肽试剂盒使用说明书中文与英文对照

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。

Specimen requirements

1.       extract as soon as possible after Specimen collection,and according to the relevant literature, and should be experiment as soon as possible after the extraction.If it can’t, specimen can be kept in -20 ℃ to preserve, Avoid repeated freeze-thaw cycles.

2.       Can’t detect the sample which contain NaN3, because NaN3 inhibits HRP active.

人血纤肽试剂盒操作步骤

1.         标准品的稀释:本试剂盒提供原倍标准品一支,用户可按照下列图表在小试管中进行稀释。

60pg/ml

5号标准品

150μl的原倍标准品加入150μl标准品稀释液

30pg/ml

4号标准品

150μl5号标准品加入150μl标准品稀释液

15pg/ml

3号标准品

150μl4号标准品加入150μl标准品稀释液

7.5pg/ml

2号标准品

150μl3号标准品加入150μl标准品稀释液

3.75pg/ml

1号标准品

150μl2号标准品加入150μl标准品稀释液

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)、标准孔、待测样品孔。在酶标包被板上标准品准确加样50μl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20倍浓缩洗涤液用蒸馏水20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:Dilute Original density Standard as follow table:


400 pg/ml


5 Standard


150μl Original density Standard+150μl Standard diluent


200 pg/ml


4 Standard


150μl 5 Standard+150μl Standard diluent


100 pg/ml


3 Standard


150μl 4 Standard+150μl Standard diluent


50 pg/ml


2 Standard


150μl 3 Standard +150μl Standard diluent


25 pg/ml


1 Standard


150μl 2 Standard +150μl Standard diluent

2. add sample:Set blank wells separately (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 at37℃.

4.Configurate liquid: 30-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.

人血纤肽试剂盒注意事项

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。

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,推荐使用排枪加样。

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。

Important notes

1.       The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag.

2.       washing buffer will Crystallization separation, it can be heated the water helps dissolve when dilute . Washing does not affect the result.

3.       add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 min, if the number of sample is much , recommend to use Volley .

4.       if the testing material content is excessively higher (The sample OD is bigger than the first standard well ),please dilute Sample (n-fold), Please diluente andmultiplied by the dilution factor.(×n×5).

5.       Closure plate membrane only limits the disposable use, to avoid cross-contamination.

6.       The substrate evade the light preservation.

7.       Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard.

8.       All samples, washing buffer and each kind of reject should according to infective material process.

9.       Do not mix reagents with those from other lots.

人血纤肽试剂盒保存条件及有效期

1.试剂盒保存:2-8

2.有效期:6个月

Storage and validity

1.Storage:  2-8℃.

2.validity: six months.

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