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原产地：中国大陆发布时间：2022/4/11 14:10:22
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本公司4月11日14时12分报道：（http://www.app17.com/c110001/products/d11333583.html）
4. 严格按照说明书中标明的时间、加液量及顺序进行温育操作。
5. 所有液体组分使用前充分摇匀。
试剂盒组成
洗板方法
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。
| 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 |
2. 自动洗板机：每孔注入洗液 350μL，浸泡 1min，洗板 5 次。操作步骤
1. 从室温平衡 20min 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；
注：标准品浓度依次为：40、20、10、5、2.5、0 ng/mL.
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
试剂的准备
的检测抗体 100μL，用封板膜封住反应孔，37℃水浴锅或恒温箱温
20×洗涤缓冲液的稀释：蒸馏水按 1：20 稀释，即 1 份的 20×洗涤
育 60min。
缓冲液加 19 份的蒸馏水。
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. 每孔加入底物 A、B 各 50μL，37℃避光孵育 15min。
7. 每孔加入终止液 50μL，15min 内，在 450nm 波长处测定各孔的OD 值。
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
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. 灵敏度：检测浓度小于 0.1 ng/mL。
3. 特异性：不与其它可溶性结构类似物交叉反应。
4. 重复性：板内变异系数小于 10%、板间变异系数小于 15%。
5. 贮藏：2-8℃，避光防潮保存。
6. 有效期：6 个月免责声明
1. 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。
2. 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。
FOR RESEARCH USE ONLY.
NOT FOR USE IN DIAGNOSTIC PROCEDURES.
Human Ureaplasma urealyticum antibody (UU-Ab)
ELISA Kit instruction
Intended use
This UU-Ab 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 UU-Ab in the sample, this UU-Ab 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 UU-Ab concentration. The concentration of UU-Ab 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 shoule be centrifugated dequately 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
Assay procedure
1. Prepare all reagent s before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.
( 20-25°C)
Materials supplied
2. Add standard: Set Standard wells, testing sample wells.
standard well.
Add standard 50μl to
| 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 |
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
Note: Standard concentration was followed by:towels.
40、20、10、5、2.5、0 ng/mL.
6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well.
Reagent preparation
Gently mix ａnd incubate for 15 minutes at 37°C. Protect from light.
20×wash solution:Dilute with Distilled or deionized water 1:20.
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
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 0.1 ng/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!
本试剂盒只能用于科学研究，不得用于医学诊断
更新时间：2024/3/6 18:05:55
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