技術文章

Technical articles

當前位置:首頁技術文章植物(Plant)脫鎂螯合酶(MDCase)ELISA檢測試劑盒

植物(Plant)脫鎂螯合酶(MDCase)ELISA檢測試劑盒

更新時間:2012-04-12點擊次數:2414

本試劑盒只能用于科學研究,不得用于醫學診斷
植物(Plant)脫鎂螯合酶(MDCase)ELISA檢測試劑盒
使用說明書
檢測原理
試劑盒采用雙抗體一步夾心法酶聯免疫吸附試驗(ELISA)。往預
先包被脫鎂螯合酶(MDCase)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,經過溫育并*洗滌。用底物TMB顯
色,TMB在過氧化物酶的催化下轉化成藍色,并在酸的作用下轉化成
zui終的黃色。顏色的深淺和樣品中的脫鎂螯合酶(MDCase)呈正相
關。用酶標儀在450nm 波長下測定吸光度(OD 值),計算樣品濃度。
樣品收集、處理及保存方法
1. 樣本不能含疊氮鈉(NaN3),因為疊氮鈉(NaN3)是辣根過氧化
物酶(HRP)的抑制劑。
2. 標本采集后盡早進行提取,提取按相關文獻進行。
3. 植物萃取液或其它相關樣本:請1000 x g離心20分鐘,取上清即
可檢測。
4. 保存:如果樣本收集后不及時檢測,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
濃縮洗滌液會有結晶,這屬于正?,F象,水浴加熱使結晶*溶解
后再使用。
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. 濃度為0 的S0 號標準品即可視為陰性對照或者空白;按照說明
書操作時樣本已經稀釋5 倍,zui終結果乘以5 才是樣本實際濃度。
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) concentration was followed by: 0,3,6,12,24,48 nmol/L
Reagent preparation
20×wash solution:Dilute with Distilled or deionized water 1:20.
Assay procedure
1. Prepare all r e a g e n 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.
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 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.

主站蜘蛛池模板: 欧美成人免费在线观看| 一级毛片免费一级直接观看| 欧美真实破苞流血在线播放| 国产强伦姧在线观看| 一本大道香焦在线视频| 欧美h版在线观看| 亚洲深深色噜噜狠狠爱网站| 色精品一区二区三区| 国产无遮挡又黄又爽网站| www.噜噜噜| 日韩美女在线观看一区| 伊人久久大香线蕉av色婷婷色| 老司机深夜福利视频| 国产精品国产福利国产秒拍| 两只大乳奶充满奶汁| 欧美18www| 亚洲成a人片77777老司机| 美女视频黄频a免费| 国产精品igao视频| www夜片内射视频日韩精品成人| 成年免费a级毛片| 亚洲av网址在线观看| 男生女生一起差差差带疼痛| 国产性生交xxxxx免费| free性欧美另类高清| 女人扒开下面让男人桶爽视频| 久久精品国产9久久综合| 玛雅视频网站在线观看免费| 国产麻豆一级在线观看| 久9re热这里精品首页| 欧美另类xxx| 免费播看30分钟大片| 青青草原综合久久大伊人| 国产成人亚洲精品播放器下载 | 在线观看国产小视频| 久久国产精品-国产精品| 欧美老妇bbbwwbbww| 又爽又刺激的视频| 黄色录像大片毛片aa| 国产肉丝袜在线观看| 一级艳片加勒比女海盗1|