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中級會員 | 第13年

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基質金屬蛋白酶/明膠酶譜法試劑盒
ELISA試劑盒
干擾物質
血清及相關類產品
生化法試劑盒
實驗代測服務項目
質控品/校準品
RIA免疫檢測
品牌生化試劑
檢測試劑盒
抗體和抗原
胰島素(Insulin)試劑盒
腫瘤壞死因子elisa試劑盒
補體蛋白試劑盒
白介素Elisa試劑盒
化學溶液
檢測試劑
商城
GIBCO培養基
染色試劑
試劑
生化指示劑
標準物質
科研用二抗
動物紅細胞懸液
生物試劑
染色液
科研細胞
生化試劑
生化實驗代測
細胞因子及重組蛋白
合成多肽

ELISA實驗英文介紹

時間:2013/5/7閱讀:3126
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ELISA (Enzyme-Linked ImmunoSorbant Assay)

ELISA實驗和ELISA試劑盒的原理介紹(英文原版)

The purpose of an ELISA is to determine if a particular protein is present in a sample and if so, how much. There are two main variations on this method: you can determine how much antibody is in a sample, or you can determine how much protein is bound by an antibody. The distinction is whether you are trying to quantify an antibody or some other protein. In this example, we will use an ELISA to determine how much of a particular antibody is present in an individuals blood.

ELISAs are performed in 96-well plates which permits high throughput results. The bottom of each well is coated with a protein to which will bind the antibody you want to measure. Whole blood is allowed to clot and the cells are centrifuged out to obtain the clear serum with antibodies (called primary antibodies). The serum is incubated in a well, and each well contains a different serum (see figure below). A positive control serum and a negative control serum would be included among the 96 samples being tested.

After some time, the serum is removed and weakly adherent antibodies are washed off with a series of buffer rinses. To detect the bound antibodies, a secondary antibody is added to each well. The secondary antibody would bind to all human antibodies and is typically produced in a rodent. Attached to the secondary antibody is an enzyme such as peroxidase or alkaline phosphatase. These enzymes can metabolize colorless substrates (sometimes called chromagens) into colored products. After an incubation period, the secondary antibody solution is removed and loosely adherent ones are washed off as before. The final step is the addition the enzyme substrate and the production of colored product in wells with secondary antibodies bound.

When the enzyme reaction is complete, the entire plate is placed into a plate reader and the optical density (i.e. the amount of colored product) is determined for each well. The amount of color produced is proportional to the amount of primary antibody bound to the proteins on the bottom of the wells.

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