计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存。低温运输 。请查阅批次 COA 获取详细规格。
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文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
本试剂盒仅供研究使用。 使用目的：本试剂盒用于测定血清，血浆及相关液体样本中乳糖(lactose)的含量。
实验原理：
本试剂盒应用酶联免疫竞争法测定标本中乳糖(lactose)水平。用纯化的乳糖(lactose)抗体包被微孔板，制成固相抗体，往包被单抗的微孔中加入乳糖(lactose)，和HRP标记的乳糖(lactose)抗原，使它们竞争结合，，经过彻底洗涤后加底物TMB显色。样本颜色的深浅和样品中的乳糖(lactose)的含量呈负相关。用酶标仪在450nm波长下测定吸光度（OD值），通过标准曲线计算样品中乳糖(lactose)的含量。
试剂盒组成：
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标本要求：
1．标本处理：
（1）水样 采集后经 -20℃反复冻融三次，再经玻璃纤维过滤后，备查
（2）组织 样品用丁醇：甲醇：水(5：25：70 V：V：V)抽提，或按相关文献提取进行，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，备查
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。
操作步骤：
1. 加样：分别设标准孔、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）、待测样品孔。在酶标包被板上标准孔中加50微升，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。
2. 加酶：每孔加入酶标试剂50μl，空白孔除外。
3. 温育：用封板膜封板后置37℃温育60分钟。
4. 配液：将30倍浓缩洗涤液用蒸馏水30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. 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.
7. 终止：每孔加终止液50μl，终止反应（此时蓝色立转黄色）。
8. 测定：以空白孔调零，450nm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。
计算：
以标准物的浓度为横坐标，OD值为纵坐标，在坐标纸上绘出标准曲线，根据样品的OD值由标准曲线查出相应的浓度；再乘以稀释倍数；或用标准物的浓度与OD值计算出标准曲线的直线回归方程式，将样品的OD值代入方程式，计算出样品浓度，再乘以稀释倍数，即为样品的实际浓度。
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。
检测范围：
2μg/L-90μg/L
This reagent kit is for research purposes only. Purpose of use: This reagent kit is used to determine the content of lactose in serum, plasma, and related liquid samples.
Experimental principle:
This kit applies enzyme-linked immunosorbent assay to determine the level of lactose in the sample. Purified lactose antibodies were coated on a microplate to produce solid-phase antibodies. Lactose was added to the microplate of the coated monoclonal antibody, along with HRP labeled lactose antigens, to compete for binding. After thorough washing, the substrate TMB was added for colorimetry. The depth of sample color is negatively correlated with the content of lactose in the sample. Measure the absorbance (OD value) at a wavelength of 450nm using an enzyme-linked immunosorbent assay (ELISA) reader, and calculate the content of lactose in the sample through a standard curve.
Kit composition:
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Specimen requirements:
1. Specimen processing:
(1) After collecting the water sample, it is repeatedly freeze-thawed three times at -20 ℃, and then filtered through glass fiber for future reference
(2) The tissue samples should be extracted using butanol: methanol: water (5:25:70 V: V: V), or extracted according to relevant literature. The experiment should be conducted as soon as possible after extraction. If the experiment cannot be conducted immediately, the specimen can be stored at -20 ℃ for future reference
2. Samples containing NaN3 cannot be detected as NaN3 inhibits the activity of horseradish peroxidase (HRP).
Operation steps:
1. Sample addition: Set up standard wells, blank wells (blank control wells do not include samples and enzyme-linked immunosorbent assay reagents, the other steps are the same), and sample wells to be tested. Add 50 microliters to the standard well on the enzyme-linked immunosorbent assay (ELISA) plate, and first add 40 diluents to the sample well to be tested μ l. Then add 10 more samples to be tested μ L (The final dilution of the sample is 5 times). Add the sample to the bottom of the enzyme-linked immunosorbent assay (ELISA) plate well, avoiding touching the well wall as much as possible. Gently shake and mix well.
2. Enzyme addition: Add 50 enzyme labeled reagents to each well μ l. Excluding blank holes.
3. Warm incubation: Seal the plate with a sealing film and incubate at 37 ℃ for 60 minutes.
4. Solution preparation: Dilute 30 times the concentrated washing solution with distilled water and set aside for later use
5. Washing: Carefully remove the sealing film, discard the liquid, shake dry, fill each well with washing solution, let it stand for 30 seconds, then discard. Repeat this process 5 times and pat dry.
6. Color development: Add color development agent A50 to each well first μ l. Add color developer B50 again μ l. Gently shake and mix well, and develop color at 37 ℃ in the dark for 15 minutes
7. Termination: Add 50% termination fluid to each hole μ l. Terminate the reaction (at this point, the blue color immediately turns yellow).
8. Measurement: Zero the blank hole and sequentially measure the absorbance (OD value) of each hole at a wavelength of 450nm. The measurement should be conducted within 15 minutes after adding the termination solution.
Calculation:
Draw a standard curve on a coordinate paper with the concentration of the standard substance as the x-axis and the OD value as the y-axis. Based on the OD value of the sample, determine the corresponding concentration from the standard curve; Multiply it by the dilution factor; Alternatively, a linear regression equation can be used to calculate the standard curve using the concentration and OD value of the standard substance. The OD value of the sample can be substituted into the equation to calculate the sample concentration, which is then multiplied by the dilution factor to obtain the actual concentration of the sample.
Notes:
1. The kit should be balanced at room temperature for 15-30 minutes before use when taken out from the cold storage environment. If the enzyme coated plate is not used up after opening, the Flat noodles should be stored in a sealed bag.
2. Concentrated detergent may precipitate crystals. When diluted, it can be heated in a water bath to aid in dissolution. Washing does not affect the results.
3. A sampler should be used for each step of sample addition, and its accuracy should be regularly calibrated to avoid experimental errors. It is best to control the sample addition time within 5 minutes. If there are a large number of specimens, it is recommended to use a firing gun for sample addition.
4. Please make a standard curve at the same time as each measurement, preferably with a re hole. If the content of the substance to be tested in the sample is too high (the OD value of the sample is greater than the OD value of the first well of the standard well), please dilute the sample diluent by a certain multiple (n times) before measurement. When calculating, please multiply the total dilution multiple (x n x 5).
5. The sealing film is only for one-time use to avoid cross contamination.
6. Please store the substrate in dark.
7. Strictly follow the instructions and determine the test results based on the reading of the enzyme-linked immunosorbent assay (ELISA) reader
8. All samples, washing liquids, and various waste should be treated as infectious substances.
9. The components of this reagent with different batch numbers shall not be mixed.
Detection range:
two μ G/L-90 μ G/L
规格
规格或纯度
96T
英文名称
Lactose Assay ELISA Kit
储存与运输
储存条件
2-8°C储存
运输条件
低温运输
技术文档
高级数据
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 H2427520 |
分析证书 | L486307 |
溶液计算器
常见问题
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