小鼠FGF-21 ELISA化学药品盒肯定是如何正常用

本化学试剂盒才能使用来科学技术深入分析，不能使用来医学专业物理诊断

小鼠成纤维细胞生长因子21（FGF-21）

ELISA查测化学制剂盒

用解释书

检查测量机理

试剂盒采用双抗体夹心法酶联免疫吸附试验（ELISA）。往预先包被小鼠成纤维细胞生长因子21（FGF-21）捕获抗体的包被微孔中，依次加入标本、标准品、HRP标记的检测抗体，经过温育并*洗涤。用底物TMB显色，TMB在过氧化物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小鼠成纤维细胞生长因子21（FGF-21）呈正相关。用酶标仪在450nm 波长下测定吸光度（OD 值），计算样品浓度。

备样抽取、工作及留存的方法

1.  血清：使用不含热原和内毒素的试管，操作过程中避免任何细胞刺激，收集血液后，3000转离心10分钟将血清和红细胞迅速小心地分离。

2.  血浆：EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。

3.  细胞上清液：3000转离心10分钟去除颗粒和聚合物。

4.  组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。

5.  保存：如果样本收集后不及时检测，请按一次用量分装，冻存于-20℃，避免反复冻融，在室温下解冻并确保样品均匀地充分解冻。

自购危险物品

1.  酶标仪（450nm）

2.  测微仪等级加样器及枪头：0.5-10uL、2-20uL、20-200uL、200-1000uL

3.  37℃控温箱

运行特别留意事宜

1.  试剂盒保存在2-8℃，使用前室温平衡60分钟。从冰箱取出的浓缩洗涤液会有结晶，这属于正常现象，水浴加热使结晶*溶解后再使用。模本在应用前也会在常温平衡量60几分钟。

2.  实验报告中不要的板条应再次放回自封袋中，密封胶（常温晾干）永久保存。

3.  预处理后的样本无需稀释，直接取10μL加样即可。

4.  须严格遵照详细说明书里表示的时候、加液量及次序开展温育运作。

5.  所有液体组分使用前充分摇匀。

实验试剂盒组成的

注：规则品氨水浓度顺序为：1600、800、400、200、100、0 pg/mL。

制剂的提前准备

 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。

洗板的方式

1.  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。

2.  全自动洗板机：每孔添加洗液350μL，泡浸1min，洗板5次。

基本操作具体步骤

1.  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℃。

2.  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；

3.  待测样本孔先加待测样本10μL，再加样本稀释液40μL；

4.  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。

5.  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）。

6.  每孔倒入底物A、B各50μL，37℃阴凉孵育15min。

7.  每孔加入终止液50μL，15min内，在450nm波长处测定各孔的OD值。

报告单诊断

 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。

 采血管盒特点

1. 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。

2.  精确度度：检测浓度小于10 pg/mL。

3.  在线检测空间：50 pg/mL - 1600 pg/mL  。                                           

4. 特情人：不与所有可可溶性组成部分接近物平行不良反应。

5.  重复性：板内变异系数小于10%、板间变异系数小于15%。

6.  贮藏：2-8℃，阴凉防尘保存图片。

7.  行之有效期限：6个月大

免责声明范文

1.   工作操作试剂盒全部理论探究便用，不得不用到理论探究工作操作或小鼠体工作操作，要不然主产地生的万事万物结果，由工作操作者承担起，本集团公司不许承担责任。

2.   严苛以规格书怎么写书运营，科学试验者触范规格书怎么写书运营，责任由科学试验者履行。

FOR RESEARCH USE ONLY.

NOT FOR USE IN DIAGNOSTIC PROCEDURES.

Mouse fibroblast growth factor-21 (FGF-21) ELISA Kit instruction

Intended use

This FGF-21 ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures. The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of FGF-21 in the sample, this FGF-21 ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus FGF-21 concentration. The concentration of FGF-21 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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

Note: Standard concentration was followed by:

1600、800、400、200、100、0 pg/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 nappear 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

1.         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.

2.         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.

3.         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.

4.         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.

5.         The sensitivity by this assay is 10 pg/mL

6.         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!

 小鼠FGF-21 ELISA化学制剂盒想必怎么样去 精准食用