无码一区二区三区久久精品|呦女1300部真实u女|人妻无码一区二区视频观看|精品乱码久久国产av大片

熱門(mén)搜索: 96T/48T植物茉莉酸甲酯(MeJA)ELISA試劑盒 96T/48T植物茉莉酸(JA)ELISA試劑盒 96T/48T植物可溶性淀粉(s-starch)ELISA試劑盒 96T/48T植物果聚糖合成酶(FBEs)ELISA試劑盒 96T/48T植物獨腳金內酯(SLs)ELISA試劑盒 96T/48T植物赤霉素3(GA3)ELISA試劑盒 96T/48T植物丙二烯氧化物合酶(AOS)ELISA試劑盒 96T/48T植物NADP蘋(píng)果酸酶(NADP-ME)ELISA試劑盒 96T/48T微生物脂肪酶(Lipase)ELISA試劑盒 96T/48T微生物胱硫醚-γ-裂解酶(CSE)ELISA試劑盒 96T/48T微生物蛋白質(zhì)水解酶(protease)ELISA試劑盒 96T/48T微生物甲酸脫氫酶(FDH)ELISA試劑盒 96T/48T微藻抗壞血酸過(guò)氧化物酶(APX)ELISA試劑盒 96T/48T微藻葉綠素(chlorophyll)ELISA試劑盒 96T/48T微藻磷酸甘油酸激酶(PGK)ELISA試劑盒 96T/48T微藻Rubisco活化酶(RCA)ELISA試劑盒

技術(shù)文章/ Technical Articles

您的位置:首頁(yè)  /  技術(shù)文章  /  人磷酸化Tau217蛋白(p-Tau217) ELISA檢測試劑盒 使用說(shuō)明書(shū)

人磷酸化Tau217蛋白(p-Tau217) ELISA檢測試劑盒 使用說(shuō)明書(shū)

更新時(shí)間:2023-09-27      瀏覽次數:735

本試劑盒只能用于科學(xué)研究,不得用于醫學(xué)診斷

Human磷酸化Tau217蛋白(p-Tau217)

ELISA檢測試劑盒

使用說(shuō)明書(shū)

檢測原理

試劑盒采用雙抗體一步夾心法酶聯(lián)免疫吸附試驗(ELISA)。往預先包磷酸化Tau217蛋白(p-Tau217)抗體的包被微孔中,依次加入標本、標準品、HRP標記的檢測抗體,經(jīng)過(guò)溫育并徹-底洗滌。用底物TMB顯色,TMB在過(guò)氧化物酶的催化下轉化成藍色,并在酸的作用下轉化成最終的黃色。顏色的深淺和樣品中的磷酸化Tau217蛋白(p-Tau217)正相關(guān)。用酶標儀在450nm 波長(cháng)下測定吸光度(OD 值),計算樣品濃度。

樣品收集、處理及保存方法

1.  血清:使用不含熱原和內毒素的試管,操作過(guò)程中避免任何細胞刺激,收集血液后,3000轉離心10分鐘將血清和紅細胞迅速小心地分離。

2.  血漿:EDTA、檸檬酸鹽或肝素抗凝。3000轉離心30分鐘取上清。

3.  細胞上清液:3000轉離心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轉離心10分鐘取上清。

5.  保存:如果樣本收集后不及時(shí)檢測,請按一次用量分裝,凍存于-20℃,避免反復凍融,在室溫下解凍并確保樣品均勻地充分解凍。

自備物品

1. 酶標儀(450nm)

2. 高精度加樣器及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL

3. 37℃恒溫箱

操作注意事項

1.  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 )有結晶,這屬于正?,F象,水浴加熱使結晶完-全溶解后再使用。

2.  實(shí)驗中不用的板條應立即放回自封袋中,密封(低溫干燥)保存。

3.  濃度為0的S0號標準品即可視為陰性對照或者空白;按照說(shuō)明書(shū)操作時(shí)樣本已經(jīng)稀釋5倍,最終結果乘以5才是樣本實(shí)際濃度。

4.  嚴格按照說(shuō)明書(shū)中標明的時(shí)間、加液量及順序進(jìn)行溫育操作。

5.  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

微孔酶標板

12孔×8條

12孔×4條

無(wú)

標準品

0.3mL*6管

0.3mL*6管

無(wú)

樣本稀釋液

6mL

3mL

無(wú)

檢測抗體-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:標準品(S0-S5)濃度依次為:0、125、250、500、1000、2000 pg/mL

試劑的準備

 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

1.  手工洗板:甩盡孔內液體,每孔加滿(mǎn)洗滌液,靜置1min后甩盡孔內液體,在吸水紙上拍干,如此洗板5次。

2.  自動(dòng)洗板機:每孔注入洗液350μL,浸泡1min,洗板5次。

操作步驟

1.  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℃。

2.  設置標準品孔和樣本孔,標準品孔各加不同濃度的標準品50μL;

3.  樣本孔先加待測樣本10μL,再加樣本稀釋液40μL;空白孔不加

4.  除空白孔外,標準品孔和樣本孔中每孔加入辣根過(guò)氧化物酶(HRP)標記的檢測抗體100μL,用封板膜封住反應孔,37℃水浴鍋或恒溫箱溫育60min。

5.  棄去液體,吸水紙上拍干,每孔加滿(mǎn)洗滌液,靜置1min,甩去洗滌液,吸水紙上拍干,如此重復洗板5次(也可用洗板機洗板)。

6.  每孔加入底物A、B各50μL,37℃避光孵育15min。

7.  每孔加入終止液50μL,15min內,在450nm波長(cháng)處測定各孔的OD值。

結果判斷

 繪制標準曲線(xiàn):在Excel工作表中,以標準品濃度作橫坐標,對應OD值作縱坐標,繪制出標準品線(xiàn)性回歸曲線(xiàn),按曲線(xiàn)方程計算各樣本濃度值。

試劑盒性能

1.  準確性:標準品線(xiàn)性回歸與預期濃度相關(guān)系數R值,大于等于0.9900。

2.  靈敏度:最-低檢測濃度小于10 pg/mL。

3.  特異性:不與其它可溶性結構類(lèi)似物交叉反應。

4.  重復性:板內、板間變異系數均小于15%。

5.  貯藏:2-8℃,避光防潮保存。

6.  有效期:6個(gè)月

免責聲明

1.   試劑盒僅供研究使用,不得用于臨床實(shí)驗或體實(shí)驗,否則所產(chǎn)生的一切后果,由實(shí)驗者承擔,本公司概不負責。

2.   嚴格按照說(shuō)明書(shū)操作,實(shí)驗者違反說(shuō)明書(shū)操作,后果由實(shí)驗者承擔。

 

FOR RESEARCH USE ONLY. 

NOT FOR USE IN DIAGNOSTIC PROCEDURES.

 

Human  tau (p-Tau217) ELISA Kit instruction

 

Intended use

This p-Tau217 ELISA kit is intended LaboHumanory 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 concentHumanion of Tau in the sample, this p-Tau217 ELISA Kit includes a set of calibHumanion standards. The calibHumanion standards are assayed at the same time as the samples and allow the opeHumanor to produce a standard curve of Optical Density versus p-Tau217 concentHumanion. The concentHumanion of p-Tau217 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 sepaHumanor 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 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 are matched for optimal performance. Use only the reagents supplied by manufacturer.

2.  Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided.

3.  Mix all reagents before using.

Remove all kit reagents from refrigeHumanor and allow them to reach room tempeHumanure ( 20-25°C)

Materials supplied

Name

96 determinations

48 determinations

Microelisa stripplate

12*8strips

12*4strips

Standard

0.3ml*6tubes

0.3ml*6tubes

 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 (S0 S5) concentHumanion was followed by:0,125,250,500,1000,2000 pg/ml

Reagent prepaHumanion

20×wash solution:Dilute with Distilled 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 to the Microelisa Stripplate.

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

3.  Add Sample: Add testing sample 10μl then add 40μl of Sample Diluent to testing sample well; Blank well doesnt add anyting.

4.  Add 10l of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C. 

5.  AspiHumane 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, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspiHumaning 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 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 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 a microtiter 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 geneHumaned by plotting the average O.D. (450 nm) obtained for each of the six standard concentHumanions on the vertical (Y) axis versus the corresponding concentHumanion 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 concentHumanion.

4. Any variation in opeHumanor, pipetting and washing technique, incubation time or tempeHumanure, 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!


微信掃一掃

郵箱:2924516602@qq.com

傳真:02166566405、19121610072

地址:上海市奉賢區金海公路6055號11幢5層

Copyright © 2024 上海佰利萊生物科技有限公司版權所有   備案號:滬ICP備2022021373號-4   技術(shù)支持:智慧城市網(wǎng)