人甲型流感（H1N1）ELISA试剂盒
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| Name | 96determinations | 48determinations |
| 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 |
Note: Standard concentration was followed by:
20、10、5、2.5、1.25、0 ng/mL.
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 appear uniform,gently tap the plate to ensure thorough mixing.8. ReadtheOpticalDensity(O.D.)at450nmusinga microtiterplatereaderwithin15minutes.Calculation of results- 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.
- 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.
- 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.
- 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.
- The sensitivity by this assay is 0.1 ng/mL.
- 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!上海瓦兰专业代理各大小医院,高校实验室,科研单位,疾控中心等机构常用的科研检测酶免
ELISA试剂盒,种属包括:人(human),大鼠(rat),小鼠(mouse),豚鼠(guinea pig),犬(dog),兔(rabbit),猪(porcine),牛(bovine),羊(goat),猴(monkey)等。
品牌主要有美国酶免(ELISA)试剂盒,美国酶免(ELISA)试剂盒,美国酶免(ELISA)试剂盒,原装/分装等。
◇质量保证：
检测用所有试剂全部都为进口试剂，适合检测包括血清、血浆、尿液、胸腹水、灌洗液、脑脊液、细胞培养上清、组织匀浆等标本，灵敏度极高，多年专业酶免服务，我们保证对所售任何产品一概负责到底，每一份实验结果都真实可靠！
◇注意事项：
在收集标本前都必须有一个完整的计划，必须清楚要检测的成份是否足够稳定。我们提倡新鲜标本尽早检测，对收集后当天就进行检测的标本，及时储存在4℃备用，如有特殊原因需要周期收集标本，请造模取材后，将标本及时分装后放在-20℃或-80℃条件下保存。因冰室与室温存在一定温差，蛋白极易降解，直接影响实验质量，所以避免反复冻融。
收集标本前必须清楚要检测的成份是否足够稳定。对收集后当天进行检测的标本，储存在4 ℃备用，如有特殊原因需要周期收集标本，将标本及时分装后放在-20 ℃或-70 ℃条件下保存。避免反复冻融。标本2 -8 ℃可保存48 小时，-20 ℃可保存1 个月。-70 度可保存6 个月。部分激素类标本需添加抑肽酶。
各检测项目酶免(ELISA)试剂盒具备,价格优惠,定货及时,送货上门.此外,我公司还提供样本代测服务。
同时公司还经营各类酶免定量(ELISA Quantikine)
试剂盒细胞因子试剂系列
具体详情请来电咨询。
