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2. 實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. 預處理后的樣本無需稀釋,直接取 10μL 加樣即可。
4. 嚴格按照說明書中標明的時間、加液量及順序進行溫育操作。
5. 所有液體組分使用前充分搖勻。
試劑盒組成
名稱 | 96 孔配置 | 48 孔配置 | 備注 |
微孔酶標板 | 12 孔×8 條 | 12 孔×4 條 | 無 |
標準品 | 0.3mL | 0.3mL | 無 |
樣本稀釋液 | 6mL | 3mL | 無 |
檢測抗體-HRP | 10mL | 5mL | 無 |
20×洗滌緩沖液 | 25mL | 15mL | 按說明書進行稀釋 |
底物A | 6mL | 3mL | 無 |
底物B | 6mL | 3mL | 無 |
終止液 | 6mL | 3mL | 無 |
封板膜 | 2 張 | 2 張 | 無 |
說明書 | 1 份 | 1 份 | 無 |
自封袋 | 1 個 | 1 個 | 無 |
注:標準品濃度依次為:20、10、5、2.5、1.25、0 ng/mL.
20×洗滌緩沖液的稀釋:蒸餾水按 1:20 稀釋,即 1 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. 待測樣本孔先加待測樣本 10μL,再加樣本稀釋液 40μL;
4. 隨后標準品孔和樣本孔中每孔加入辣根過氧化物酶(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 波長處測定各孔的 OD 值。
結果判斷
繪制標準曲線:在 Excel 工作表中,以標準品濃度作橫坐標,對應OD 值作縱坐標,繪制出標準品線性回歸曲線,按曲線方程計算各樣本濃度值。
試劑盒性能
1. 準確性:標準品線性回歸與預期濃度相關系數 R 值,大于等于0.9900。
2. 靈敏度:檢測濃度小于 0.1 ng/mL。
3. 特異性:不與其它可溶性結構類似物交叉反應。
4. 重復性:板內變異系數小于 10%、板間變異系數小于 15%。
5. 貯藏:2-8℃,避光防潮保存。
6. 有效期:6 個月
免責聲明
1. 試劑盒僅供研究使用,不得用于臨床實驗或人體實驗,否則所產生的一切后果,由實驗者承擔,本公司概不負責。
2. 嚴格按照說明書操作,實驗者違反說明書操作,后果由實驗者承擔。
FOR RESEARCH USE ONLY.
NOT FOR USE IN DIAGNOSTIC PROCEDURES.
Human Thyroid stimulating antibody (TSAb) ELISA Kit instruction
Intended use
This TSAb 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 TSAb in the sample, this TSAb 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 TSAb concentration. The concentration of TSAb in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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 shoule be centrifugated dequately and no hemolysis or granule was allowed.
1. Standard microplate reader(450nm)
2. Precision pipettes and Disposable pipette tips.
3. 37 ℃ incubator
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 refrigerator and allow them to reach room temperature ( 20-25°C)
Name | 96 determinations | 48 determinations |
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.
20×wash solution:Dilute with Distilled or deionized water 1:20.
1. Prepare all r eagen t s 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 sample 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 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 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 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.
本試劑盒只能用于科學研究,不得用于醫學診斷
更新時間:2024/5/24 11:45:35
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