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：最低检测浓度小于0.1mmol/L。

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.

HumanLow Density Lipoprotein(LDL) ELISA Kit instruction

Intended use

This LDL 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 LDL in the sample, this LDL 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 LDL concentration. The concentration of LDL in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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

Note: Standard concentration was followed by:

8、4、2、1、0.5、0 mmol/L.

Reagent preparation

20×wash solution:Dilute with Distilled or deionized water 1:20.

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 sample 10μl Then add sample diluent 40μl to testing sample well; Blank well doesn’t add anyting.

4. Add100μlofHRP-conjugate reagent to 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