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：低检测浓度小于10 pg/mL。

3.  检测范围：10.0-2400 pg/mL

4.  特异性：不与其它可溶性结构类似物交叉反应。

5.  重复性：板内、板间变异系数均小于15%。

6.  贮藏：2-8℃，避光防潮保存。

7.  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.

Rat CXCL11 ELISA Kit instruction

Intended use

ThisCXCL11 ELISA kit is intended Laboratory for Research use only ａnd is not for use in diagnostic or therapeutic procedures.The S 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 ofCXCL11 in the sample, thisCXCL11 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 versusCXCL11 concentration. The concentration ofCXCL11 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 for10 minutes at approximately3000×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 for30 minutes at3000×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 arematched for optimal performance. Use onlythe reagents supplied bymanufacturer.

2.  Do not remove microplate from thestorage bag until needed. Unused strips should bestored at 2-8°C intheir pouch with the desiccant provided.

3. Mix all reagents before using.

Remove all kit reagents from refrigerator ａnd allow them toreach room temperature ( 20-25°C)

Materials supplied

Note:Standard (S0 →S5) concentration was followedby:0,150,300,600,1200,2400 pg/ml

Reagent preparation

20×wash solution:Dilute withDistilled 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 totheMicroelisa Stripplate.

2. Addstandard:Set Standard wells,testing sample wells.Add standard 50μl to standard well.

3.  AddSample: Add testing sample 10μlthen addSample 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 processfour times for a total of fivewashes. Wash by filling each well with WashSolution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to goodperformance. After the last wash, remove any remaining WashSolution 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 mixandincubate for 15 minutes at37°C.Protect from light.

7. Add 50μlS 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 amicrotiter 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!