人（Human）葉(ye)酸(suan)（folic acid）ELISA檢(jian)測(ce)試(shi)劑(ji)盒

本試劑盒只(zhi)能(neng)用於科學(xue)研究，不(bu)得(de)用(yong)於醫(yi)學診(zhen)斷
人（Human）葉(ye)酸(suan)（folic acid）ELISA檢(jian)測(ce)試(shi)劑(ji)盒
使(shi)用(yong)說(shuo)明書(shu)
檢(jian)測(ce)原(yuan)理(li)
試(shi)劑盒采(cai)用(yong)雙(shuang)抗體壹(yi)步(bu)夾(jia)心法(fa)酶(mei)聯免(mian)疫(yi)吸附(fu)試(shi)驗（ELISA）。往(wang)預(yu)
先(xian)包被(bei)葉(ye)酸(suan)（folic acid）抗(kang)體(ti)的(de)包被(bei)微孔(kong)中(zhong)，依次加入(ru)標本、標準品(pin)、
HRP標記的(de)檢(jian)測(ce)抗(kang)體(ti)，經(jing)過(guo)溫育(yu)並*洗滌。用(yong)底(di)物(wu)TMB顯色(se)，TMB
在(zai)過(guo)氧(yang)化(hua)物(wu)酶(mei)的(de)催化(hua)下(xia)轉化(hua)成(cheng)藍色，並在酸(suan)的(de)作(zuo)用(yong)下轉(zhuan)化(hua)成(cheng)zui終的(de)
黃(huang)色(se)。顏色(se)的(de)深(shen)淺(qian)和樣(yang)品(pin)中的(de)葉(ye)酸(suan)（folic acid）呈正(zheng)相關。用酶(mei)標儀
在450nm 波(bo)長下(xia)測(ce)定(ding)吸光度(du)（OD 值），計算(suan)樣(yang)品(pin)濃(nong)度(du)。
樣(yang)品(pin)收(shou)集、處(chu)理(li)及(ji)保(bao)存(cun)方(fang)法(fa)
1. 血(xue)清：使(shi)用不(bu)含熱原(yuan)和(he)內毒素(su)的(de)試(shi)管(guan)，操作(zuo)過(guo)程(cheng)中(zhong)避(bi)免(mian)任(ren)何(he)細(xi)胞(bao)
刺激，收(shou)集血(xue)液後，3000 轉離(li)心(xin)10 分(fen)鐘(zhong)將(jiang)血(xue)清和(he)紅細(xi)胞迅速小心地
分(fen)離(li)。
2. 血(xue)漿(jiang)：EDTA、檸(ning)檬(meng)酸(suan)鹽或肝(gan)素(su)抗凝(ning)。3000 轉離(li)心(xin)30 分(fen)鐘(zhong)取上(shang)清(qing)。
3. 細胞(bao)上清液：3000 轉離(li)心(xin)10 分(fen)鐘(zhong)去除(chu)顆粒(li)和聚(ju)合(he)物(wu)。
4. 組織(zhi)勻(yun)漿(jiang)：將(jiang)組織(zhi)加入(ru)適(shi)量生(sheng)理(li)鹽水搗碎。3000 轉離(li)心(xin)10 分(fen)鐘(zhong)
取上(shang)清(qing)。
5. 保(bao)存(cun)：如果(guo)樣(yang)本收(shou)集後不及(ji)時(shi)檢(jian)測(ce)，請(qing)按(an)壹次(ci)用量(liang)分(fen)裝，凍(dong)存於
-20℃，避免(mian)反復(fu)凍(dong)融(rong)，在室(shi)溫下解凍(dong)並確(que)保(bao)樣(yang)品(pin)均勻(yun)地(di)充分(fen)解(jie)凍(dong)。
自備物(wu)品(pin)
1. 酶(mei)標儀（450nm）
2. 高精度(du)加樣(yang)器及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL
3. 37℃恒(heng)溫箱(xiang)
操作註(zhu)意(yi)事(shi)項
1. 試(shi)劑盒保(bao)存(cun)在(zai)2-8℃，使(shi)用前室(shi)溫平衡20 分(fen)鐘(zhong)。從(cong)冰箱(xiang)取出(chu)的(de)
濃(nong)縮洗(xi)滌(di)液(ye)會有(you)結(jie)晶(jing)，這(zhe)屬於正(zheng)常現象，水(shui)浴加熱使(shi)結(jie)晶(jing)*溶解
後再使(shi)用(yong)。
2. 實(shi)驗中不(bu)用的(de)板條應立即放回自封袋中(zhong)，密封（低(di)溫幹燥(zao)）保(bao)存(cun)。
3. 濃(nong)度(du)為(wei)0 的(de)S0 號標準品(pin)即可視為(wei)陰(yin)性(xing)對(dui)照或者空白；按(an)照說明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:0,1.5,3,6,12,24 ng/ml
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 esting 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