亚洲无码在线播放,欧美性爱一级在线视频,最新国产在线拍揄自揄视频,国产福利在线观看不卡视频,亚洲中文字幕aⅴ无码

<wbr id="au4ne"></wbr>
    1. <source id="au4ne"></source>

      大鼠活性氧簇(ROS)ELISA檢測(cè)試劑盒

      發(fā)布時(shí)間:2024-09-20
      本試劑盒只能用于科學(xué)研究,不得用于醫(yī)學(xué)診斷
      大鼠活性氧簇(ros)elisa檢測(cè)試劑盒
      使用說(shuō)明書(shū)
      檢測(cè)原理
      試劑盒采用雙抗體夾心法酶聯(lián)免疫吸附試驗(yàn)(elisa)。往預(yù)先包被大鼠活性氧簇(ros)捕獲抗體的包被微孔中,依次加入標(biāo)本、標(biāo)準(zhǔn)品、hrp標(biāo)記的檢測(cè)抗體,經(jīng)過(guò)溫育并*洗滌。用底物tmb顯色,tmb在過(guò)氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成終的黃色。顏色的深淺和樣品中的大鼠活性氧簇(ros)呈正相關(guān)。用酶標(biāo)儀在450nm 波長(zhǎng)下測(cè)定吸光度(od 值),計(jì)算樣品濃度。
      樣品收集、處理及保存方法
      1. 血清:使用不含熱原和內(nèi)毒素的試管,操作過(guò)程中避免任何細(xì)胞刺激,收集血液后,3000轉(zhuǎn)離心10分鐘將血清和紅細(xì)胞迅速小心地分離。
      2. 血漿:edta、檸檬酸鹽或肝素抗凝。3000轉(zhuǎn)離心30分鐘取上清。
      3. 細(xì)胞上清液:3000轉(zhuǎn)離心10分鐘去除顆粒和聚合物。
      4. 組織勻漿:將組織加入適量生理鹽水搗碎。3000轉(zhuǎn)離心10分鐘取上清。
      5. 保存:如果樣本收集后不及時(shí)檢測(cè),請(qǐng)按一次用量分裝,凍存于-20℃,避免反復(fù)凍融,在室溫下解凍并確保樣品均勻地充分解凍。
      自備物品
      酶標(biāo)儀(450nm)高精度加樣器及槍頭:0.5-10ul、2-20ul、20-200ul、200-1000ul37℃恒溫箱操作注意事項(xiàng)
      試劑盒保存在2-8℃,使用前室溫平衡20分鐘。從冰箱取出的濃縮洗滌液會(huì)有結(jié)晶,這屬于正?,F(xiàn)象,水浴加熱使結(jié)晶*溶解后再使用。實(shí)驗(yàn)中不用的板條應(yīng)立即放回自封袋中,密封(低溫干燥)保存。預(yù)處理后的樣本無(wú)需稀釋?zhuān)苯尤?0μl加樣即可。嚴(yán)格按照說(shuō)明書(shū)中標(biāo)明的時(shí)間、加液量及順序進(jìn)行溫育操作。所有液體組分使用前充分搖勻。試劑盒組成
      名稱(chēng)
      96孔配置
      48孔配置
      備注
      微孔酶標(biāo)板
      12孔×8條
      12孔×4條
      無(wú)
      標(biāo)準(zhǔn)品
      0.3ml
      0.3ml
      無(wú)
      *
      6ml
      3ml
      無(wú)
      檢測(cè)抗體-hrp
      10ml
      5ml
      無(wú)
      20×洗滌緩沖液
      25ml
      15ml
      按說(shuō)明書(shū)進(jìn)行稀釋
      底物a
      6ml
      3ml
      無(wú)
      底物b
      6ml
      3ml
      無(wú)
      終止液
      6ml
      3ml
      無(wú)
      封板膜
      2張
      2張
      無(wú)
      說(shuō)明書(shū)
      1份
      1份
      無(wú)
      自封袋
      1個(gè)
      1個(gè)
      無(wú)
      注:標(biāo)準(zhǔn)品濃度依次為:480、240、120、60、30、0 u/ml.
      試劑的準(zhǔn)備
      20×洗滌緩沖液的稀釋?zhuān)赫麴s水按1:20稀釋?zhuān)?份的20×洗滌緩沖液加19份的蒸餾水。
      洗板方法
      手工洗板:甩盡孔內(nèi)液體,每孔加滿洗滌液,靜置1min后甩盡孔內(nèi)液體,在吸水紙上拍干,如此洗板5次。自動(dòng)洗板機(jī):每孔注入洗液350μl,浸泡1min,洗板5次。操作步驟
      從室溫平衡20min后的鋁箔袋中取出所需板條,剩余板條用自封袋密封放回4℃。設(shè)置標(biāo)準(zhǔn)品孔和樣本孔,標(biāo)準(zhǔn)品孔各加不同濃度的標(biāo)準(zhǔn)品50μl;待測(cè)樣本孔先加待測(cè)樣本10μl,再加*40μl;隨后標(biāo)準(zhǔn)品孔和樣本孔中每孔加入辣根過(guò)氧化物酶(hrp)標(biāo)記的檢測(cè)抗體100μl,用封板膜封住反應(yīng)孔,37℃水浴鍋或恒溫箱溫育60min。棄去液體,吸水紙上拍干,每孔加滿洗滌液,靜置1min,甩去洗滌液,吸水紙上拍干,如此重復(fù)洗板5次(也可用洗板機(jī)洗板)。每孔加入底物a、b各50μl,37℃避光孵育15min。每孔加入終止液50μl,15min內(nèi),在450nm波長(zhǎng)處測(cè)定各孔的od值。結(jié)果判斷
      繪制標(biāo)準(zhǔn)曲線:在excel工作表中,以標(biāo)準(zhǔn)品濃度作橫坐標(biāo),對(duì)應(yīng)od值作縱坐標(biāo),繪制出標(biāo)準(zhǔn)品線性回歸曲線,按曲線方程計(jì)算各樣本濃度值。
      試劑盒性能
      準(zhǔn)確性:標(biāo)準(zhǔn)品線性回歸與預(yù)期濃度相關(guān)系數(shù)r值,大于等于0.9900。靈敏度:低檢測(cè)濃度小于1.0u/ml。特異性:不與其它可溶性結(jié)構(gòu)類(lèi)似物交叉反應(yīng)。重復(fù)性:板內(nèi)變異系數(shù)小于10%、板間變異系數(shù)小于15%。貯藏:2-8℃,避光防潮保存。有效期:6個(gè)月免責(zé)聲明
      試劑盒僅供研究使用,不得用于臨床實(shí)驗(yàn)或人體實(shí)驗(yàn),否則所產(chǎn)生的一切后果,由實(shí)驗(yàn)者承擔(dān),本公司概不負(fù)責(zé)。嚴(yán)格按照說(shuō)明書(shū)操作,實(shí)驗(yàn)者違反說(shuō)明書(shū)操作,后果由實(shí)驗(yàn)者承擔(dān)。
      for research use only.
      not for use in diagnostic procedures.
      ratreactive oxygen species(ros) elisa kit instruction
      intended use
      this roselisa 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 rosin the sample, this roselisa 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 rosconcentration. the concentration of rosin 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 30minutes 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. donotsubstitutereagentsfromone kitto another.standard, conjugateandmicroplates are matchedfor optimal performance.useonly thereagentssuppliedby manufacturer.
      2. donotremovemicroplatefromthe storage baguntilneeded.unusedstripsshouldbe stored at2-8°cin their pouchwiththe desiccantprovided.
      3. mix all reagents before using.
      remove allkitreagentsfromrefrigerator and allowthemto reachroomtemperature( 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:
      480、240、120、60、30、0 u/ml.
      reagent preparation
      20×wash solution:dilute with distilled or deionized water1:20.
      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 sample10μl then add sample diluent 40μl to testing sample well; blank welldoesn’t add anyting.
      4. add100μlofhrp-conjugate reagentto each well,cover with anadhesive stripandincubatefor60 minutesat37°c.
      5. aspirate each well and wash, repeating the process fourtimes 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 solutionby 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 is1.0 u/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!
      上一個(gè):滇紅茶的泡法
      下一個(gè):木材烘干設(shè)備的技術(shù)知識(shí)

      榮耀6手機(jī)怎么拆機(jī),華為榮耀6怎么拆后蓋
      新款 醫(yī)療器械負(fù)壓測(cè)試儀 YBB00052005 測(cè)試原理
      彩色無(wú)紙記錄儀,溫度無(wú)紙記錄儀?(迅鵬)WPR80A
      工程建設(shè)項(xiàng)目前期工作流程
      數(shù)控加工中心電氣柜制造技術(shù)研究
      茶籽粉振動(dòng)篩型號(hào)和價(jià)格
      下面講一講四柱液壓機(jī)的詳細(xì)裝配過(guò)程
      閥門(mén)的特種鑄造于基本工藝
      給移動(dòng)硬盤(pán)分區(qū)工具,移動(dòng)硬盤(pán)分區(qū)要有什么工具
      茶的功用