ELISA小常識(shí)：

在目前的以HRP作為標(biāo)記酶的商品ELISA試劑盒中，如以TMB為底物，則提供的底物為A和B兩瓶應(yīng)用液；如以O(shè)PD為底物，則試劑盒提供OPD 片劑或粉劑，臨用前配制。一般商品試劑盒顯色反應(yīng)條件為37℃或室溫反應(yīng)15~30min。從理論上說(shuō)，37℃30min才可以使HRP的底物催化反應(yīng)*，盡管在zui初的10min內(nèi)，絕大部分催化反應(yīng)即可完成。因此，為使弱陽(yáng)性樣本孔能有充分的顯色，建議在37℃下反應(yīng)25~30min后，終止反應(yīng)比色測(cè)定。

產(chǎn)品說(shuō)明
eisa試劑盒:ELISA:試劑盒:酶聯(lián)免疫試劑盒:KIT

保存條件及有效期：
1.試劑盒保存：；2-8℃。
2.有效期：6個(gè)月

適用標(biāo)本:血浩、血槳、細(xì)胞焙養(yǎng)上清液、組織勻槳等

主要用途：科研檢測(cè)

原理:雙抗夾心法

試劑盒組成
1.標(biāo)準(zhǔn)品
2.標(biāo)準(zhǔn)品稀釋濃
3.酶標(biāo)包被板
4.鏈霉親和素/HRP
5.20倍濃縮洗滌濃
6.顯色劑A液
7.顯色劑B液
8.終止液等

需要而未提供的試劑和器材
1.37'C恒溫箱。
2.標(biāo)準(zhǔn)規(guī)格酶標(biāo)儀。
3.精密移液器及一次性吸頭
4.蒸餾水
5一次性試管
6.吸水紙

注意事項(xiàng)
1.從2-8'C取出的試劑盒，在開(kāi)啟試劑盒之前要室溫平衡至少30分鐘.酶標(biāo)包被板開(kāi)封后如未用完，板條應(yīng)裝入密封袋中保存.
2.各步加樣均應(yīng)使用加樣器，并經(jīng)常校對(duì)其準(zhǔn)確性，以避免試驗(yàn)誤差.
3.嚴(yán)格按照說(shuō)明書(shū)的操作進(jìn)行，試驗(yàn)結(jié)果判定必須以酶標(biāo)儀讀數(shù)為準(zhǔn).
4.為避免交叉污染，要避免重復(fù)使用手中的吸頭和封板膜.
5.不用的其它試劑應(yīng)包裝好或蓋好.不同批號(hào)的試劑不要混用.保質(zhì)前使用.
6.底物B對(duì)光敏感，避免長(zhǎng)時(shí)間暴露于光下。

標(biāo)本要求
1.不能檢測(cè)含NaN3助樣品，因NaN3抑制辣根過(guò)氧化物酶的〔HRP）活性。
2.標(biāo)本采集后盡早進(jìn)行提取，提取按相關(guān)文獻(xiàn)進(jìn)行，提取后應(yīng)盡塊進(jìn)行實(shí)驗(yàn).若不能馬上進(jìn)行試驗(yàn)，可將標(biāo)本放于-20'C保存，但應(yīng)避免反復(fù)凍融

樣品收集、處理及保存方法:
1、血清……操作過(guò)程中避免任何細(xì)胞刺激.使用不含熱原和內(nèi)毒素的試管.收集血濃后，1000xg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、肝素血漿可用于檢測(cè).1000xg離心30分鐘去除顆粒.
3.細(xì)胞上清液......1000 xg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揭碎.1000xg離心10分鐘，取上清液.
5.保存……如果樣品不立即使用，應(yīng)將其分成小部分-70'C保存，避免反復(fù)冷凍.盡可能的不要使用溶血或高血脂血.如果血清中大量顆粒，檢測(cè)前先離心或過(guò)濾。不要在37'C或更高的溫度加熱解凍。應(yīng)在室溫下解凍并確保樣品均勻地充分解凍。

具體詳見(jiàn)說(shuō)明書(shū)

Product Description
eisa kits: ELISA,: kits: ELISA kits: KIT

Storage conditions and duration of:
A kit to save:; 2-8 ° C.
Duration: 6 months

Applies to specimens: Hao of blood, plasma, cell baking raise the supernatant, the homogenate

Main purposes: scientific detection

Principle: the double-antibody sandwich method

Kit Contents
（1） standard
（2） Standard dilution of concentrated
3 ELISA coated plate
（4） streptavidin biotin / HRP
5.20 times concentrated washed concentrated
6. Color reagent A solution
7 color reagent B solution
8 Stop Solution

Without providing reagents and equipment
1.37 'C incubator.
Standard specifications microplate reader.
（3） precision pipette and the disposable tip
4. Distilled water
5 one-time test-tube
6. Absorbent paper

Note
（1） from 2-8'C remove the kit at room temperature before you open the kit balance of at least 30 minutes. ELISA plate coated plate after opening if not used, the lath should be stored in Sealed bag.
Each step of sample should be used pipette, and often proof of its accuracy, to avoid experimental error.
Strictly in accordance with the instructions of the operation, determine the test results to the microtiter plate reader as a standard.
In order to avoid cross-contamination, to avoid repeated use of the hands of the tip and the sealing plate membrane.
Without the other reagents should be packaged or covered. Do not mix different batches of reagents. Shelf life before use.
6. Substrate B is light sensitive, avoid prolonged exposure to light.

Specimen requirements
Not detect samples containing NaN3 help because of NaN3 inhibition of horseradish peroxidase [HRP） activity.
2 specimen collection as soon as possible extraction, extraction by the relevant literature, due the blck after the extraction experiment. If it can not test the specimens were stored at-20'C, but should avoid repeated freezing and thawing

Sample collection, processing and preservation methods:
Serum. Operation to avoid any cell stimulation. Pyrogen and endotoxin-free test tube to collect the blood concentration, 1000xg centrifugation for 10 minutes in serum and red blood cells rapidly and carefully isolated.
.1000 Xg for 30 minutes centrifugation, plasma ...... of EDTA. Citrate and heparin plasma can be used to detect the removal of particles.
Supernatant. 1,000 xg for centrifugal 10 minutes to remove particles and polymer.
Tissue homogenates. Will be organized by adding the appropriate amount of saline exposing broken .1000 xg for centrifuged for 10 minutes, and the supernatant.
Save ... If the sample is not used immediay, it should be divided into small part-70'C to save, to avoid repeated freezing as much as possible do not use hemolyzed or high lipid blood serum, a large number of particles, centrifugation or filtration prior to testing . Do not heated at 37'C or higher temperature to thaw. Should be thawed at room temperature and to ensure sample homogeneity fully thawed.