Parathyroid Hormone-Related Protein,PTH-RP,1-34,Human) ELISA试剂盒价格,详情介绍-960化工网

Parathyroid Hormone-Related Protein,PTH-RP,1-34,Human) ELISA试剂盒

价格：￥3200

品牌： wksubio

产地：国内

最新更新日期：2021-08-24 15:58

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上海瓦兰生物科技有限公司

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公司：上海瓦兰生物科技有限公司

联系人：陈云

联系电话： 15601655801 021-54652189

联系邮箱：walanbio@sina.com

QQ： 2530047176

产品属性

供应商

瓦兰生物

检测限

不限

检测方法

酶联免疫法

应用

科研单位

适应物种

不限

标记物	Parathyroid Hormone-Related Protein,PTH-RP,1-34,Human

样本

液体

数量

大量

规格

96T

产品说明

Parathyroid Hormone-Related Protein,PTH-RP,1-34,Human ELISA试剂盒

Samplecollection and storages
Serum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cycles
Plasma- Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 30 minutes at 3000×g at 2-8℃ within 30 minutes of collection. Store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.
Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.
Note: The samples should be centrifugated adequately and no hemolysis or granule was allowed.
Materials required but not supplied
1. Standard microplate reader(450nm)
2. Precision pipettes and Disposable pipette tips.
3. 37 ℃ incubator
Precautions
1. Donot substitutereagentsfromone kit to another.Standard, conjugateandmicroplates are matchedfor optimal performance. Useonly thereagentssuppliedby manufacturer.
2. Donot removemicroplatefrom the storage baguntilneeded. Unusedstripsshouldbe stored at2-8°Cin their pouchwith the desiccantprovided.
3. Mix all reagents before using.
Remove allkit reagentsfrom refrigerator and allow them to reachroom temperature( 20-25°C)
Materials supplied

Name	96determinations	48determinations
Microelisa stripplate	12*8strips	12*4strips
Standard	0.3ml	0.3ml
Sample diluent	6.0ml	3.0ml
HRP-Conjugate reagent	10.0ml	5.0ml
20X Wash solution	25ml	15ml
Chromogen Solution A	6.0ml	3.0ml
Chromogen Solution B	6.0ml	3.0ml
Stop Solution	6.0ml	3.0ml
Closure plate membrane	2	2
User manual	1	1
Sealed bags	1	1

Note: Standard concentration was followed by:
8、4、2、1、0.5、0 ng/ml.
Reagent preparation
20×wash solution:Dilute with Distilled or deionized water1:20.
Assay procedure
1. Prepare allreagentsbeforestartingassayprocedure. ItisrecommendedthatallStandardsand Samplesbe addedin duplicateto the MicroelisaStripplate.
2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.
3. Add Sample: Add testing sample10μl Then add sample diluent 40μl to testing sample well; Blank welldoesn’t add anyting.
4. Add100μlofHRP-conjugate reagentto each well,cover with an adhesive stripandincubatefor60 minutes at37°C.
5. Aspirate each well and wash, repeating the process four times for a total of five washes.Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifolddispenseror autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating ordecanting. Invert the plate and blot it against clean paper towels.
6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.
7. Add 50μl Stop Solution to each well. The color in the wells should change from blue toyellow. If the color in the wells is green or the color change does not

appear uniform,gently tap the plate to ensure thorough mixing.
8. ReadtheOpticalDensity(O.D.)at450nmusinga microtiterplatereaderwithin15minutes.
Calculation of results

This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis versus the corresponding concentration on the horizontal (X) axis.
First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the zero standard before result interpretation. Construct the standard curve using graph paper or statistical software.
To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration.
Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.
The sensitivity by this assay is0.1 ng/ml.
Standard curve

Storage： 2-8℃.
validity：six months.

FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS!PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!

试剂盒组成

名称	96孔配置	48孔配置	备注
微孔酶标板	12孔×8条	12孔×4条	无
标准品	0.3mL	0.3mL	无
样本稀释液	6mL	3mL	无
检测抗体-HRP	10mL	5mL	无
20×洗涤缓冲液	25mL	15mL	按说明书进行稀释
底物A	6mL	3mL	无
底物B	6mL	3mL	无
终止液	6mL	3mL	无
封板膜	2张	2张	无
说明书	1份	1份	无
自封袋	1个	1个	无

注：标准品浓度依次为：20、10、5、2.5、1.25、0 ng/mL.
试剂的准备
20×洗涤缓冲液的稀释：蒸馏水按1：20稀释，即1份的20×洗涤缓冲液加19份的蒸馏水。
洗板方法

手工洗板：甩尽孔内液体，每孔加满洗涤液，静置1min后甩尽孔内液体，在吸水纸上拍干，如此洗板5次。
自动洗板机：每孔注入洗液350μL，浸泡1min，洗板5次。

操作步骤

从室温平衡60min后的铝箔袋中取出所需板条，剩余板条用自封袋密封放回4℃。
设置标准品孔和样本孔，标准品孔各加不同浓度的标准品50μL；
待测样本孔先加待测样本10μL，再加样本稀释液40μL；
随后标准品孔和样本孔中每孔加入辣根过氧化物酶（HRP）标记的检测抗体100μL，用封板膜封住反应孔，37℃水浴锅或恒温箱温育60min。
弃去液体，吸水纸上拍干，每孔加满洗涤液，静置1min，甩去洗涤液，吸水纸上拍干，如此重复洗板5次（也可用洗板机洗板）。
每孔加入底物A、B各50μL，37℃避光孵育15min。
每孔加入终止液50μL，15min内，在450nm波长处测定各孔的OD值。

结果判断
绘制标准曲线：在Excel工作表中，以标准品浓度作横坐标，对应OD值作纵坐标，绘制出标准品线性回归曲线，按曲线方程计算各样本浓度值。

试剂盒性能

准确性：标准品线性回归与预期浓度相关系数R值，大于等于0.9900。
灵敏度：最低检测浓度小于0.1ng/mL。
特异性：不与其它可溶性结构类似物交叉反应。
重复性：板内变异系数小于10%、板间变异系数小于15%。
贮藏：2-8℃，避光防潮保存。
有效期：6个月

免责声明

试剂盒仅供研究使用，不得用于临床实验或人体实验，否则所产生的一切后果，由实验者承担，本公司概不负责。
严格按照说明书操作，实验者违反说明书操作，后果由实验者承担。

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