人胃蛋白酶 ( PG ) ELISA Kit价格_品牌:wksubio-丁香通官网

样本：液体

标记物：PG

适应物种：不限

应用：科研单位

检测方法：酶联免疫法

检测限：不限

供应商：瓦兰生物

库存：大量

规格：96T

人胃蛋白酶(PG) ELISA Kit

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：8、4、2、1、0.5、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0.1 ng/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。

◇服务内容：凡购买本司目录任何一种检测试剂盒（酶联免疫、放射免疫），您只需将需要检测的动物（Human,Rat,Mouse,Rabbit,Pig,Monkey ……）种类和检测指标（介素类、因子类）及标本数量（48T/96T）通知我司业务人员即可。在接到客户标本当日起，一周内将检测报告交到客户手中！ ◇质量保证：检测用所有试剂全部都为进口试剂，适合检测包括血清、血浆、尿液、胸腹水、灌洗液、脑脊液、细胞培养上清、组织匀浆等标本，灵敏度极高，多年专业酶免服务，我们保证对所售任何产品一概负责到底，每一份实验结果都真实可靠！ ◇注意事项：在收集标本前都必须有一个完整的计划，必须清楚要检测的成份是否足够稳定。我们提倡新鲜标本尽早检测，对收集后当天就进行检测的标本，及时储存在4℃备用，如有特殊原因需要周期收集标本，请造模取材后，将标本及时分装后放在-20℃或-70℃条件下保存。因冰室与室温存在一定温差，蛋白极易降解，直接影响实验质量，所以避免反复冻融。代测放免标本的客户取材前须向我司销售人员索要说明书，具体操作注意事项请与我司技术人员沟通。 ◇液体类标本：包括血清、血浆、尿液、胸腹水、脑脊液、细胞培养上清等。 ◇血清：室温血液自然凝固10-20分钟后，离心20分钟左右（2000-3000转/分）。仔细收集上清。保存过程中如有沉淀形成，应再次离心。 ◇血浆：应根据标本的要求选择EDTA、柠檬酸钠或肝素作为抗凝剂，混合10-20分钟后，离心20分钟左右（2000-3000转/分）。仔细收集上清。保存过程中如有沉淀形成，应再次离心。 ◇尿液：用无菌管收集。离心20分钟左右（2000-3000转/分）。仔细收集上清。保存过程中如有沉淀形成，应再次离心。胸腹水、脑脊液参照此实行。 ◇细胞培养上清：检测分泌性的成份时，用无菌管收集。离心20分钟左右（2000-3000转/分）。仔细收集上清。检测细胞内的成份时，用PBS（PH7.2-7.4）稀释细胞悬液，细胞浓度达到100万/ml左右。通过反复冻融，以使细胞破坏并放出细胞内成份。离心20分钟左右（2000-3000转/分）。仔细收集上清。保存过程中如有沉淀形成，应再次离心。 ◇组织标本：切割标本后，称取重量。加入一定量的PBS，PH7.4。用液氮迅速冷冻保存备用。标本融化后仍然保持2-8℃的温度。加入一定量的PBS（PH7.4），用手工或匀浆器将标本匀浆充分。离心20分钟左右（2000-3000转/分）。仔细收集上清。分装后一份待检测，其余冷冻备用。郑重声明：请客户在取材前尽量先与我公司销售技术人员沟通，前期的取材直接影响到您的实验结果。我们有成套的售前售后服务，凡不是在我公司购买的ELISA产品，恕我公司概不负责售后服务。

Sample collection 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. Do not substitute reagents from one kit to another. Standard, conjugate and microplates are matched for optimal performance. Use only the reagents supplied by manufacturer.
2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided.
3. Mix all reagents before using.
Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)
Materials supplied

Name	96 determinations	48 determinations
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:
20、10、5、2.5、1.25、0 ng/mL.
Reagent preparation
20×wash solution:Dilute with Distilled or deionized water 1:20.
Assay procedure
1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.
2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.
3. Add Sample: Add testing sample 10μl Then add sample diluent 40μl to testing sample well; Blank well doesn’t add anyting.
4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°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, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. 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 to yellow. 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. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.
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 is 0.1 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!