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產品分類什么是elisa試劑盒?要購買什么樣的elisa試劑盒?這個試劑盒能夠符合實驗的要求么?檢測下來的準確度怎么樣?檢測的數據用來發文章會得到認可么?這是很多老師在購買elisa試劑盒前往往會考慮的問題。
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elisa試劑盒的抉擇 請認準信帆生物
What is ELISA? Enzyme-linked immunosorbent assay (ELISA)
Enzyme-linked immunosorbent assay (ELISA), also known as an enzyme immunoassay (EIA), is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality-control check in various industries,such as ELISA application in food industry. In simple terms, in ELISA, an unknown amount of antigen is affixed to a surface, and then a specific antibody is applied over the surface so that it can bind to the antigen. This antibody is linked to an enzyme, and in the final step a substance is added that the enzyme can convert to some detectable signal, most commonly a colour change in a chemical substrate.
酶聯免疫吸附試驗(ELISA),也稱為酶免疫測定法(EIA),是一種生化技術,主要用于免疫學檢測樣品中的抗體或抗原的存在。酶聯免疫吸附試驗已作為醫學和植物病理學的診斷工具,并已在食品工業中應用,如酶聯免疫吸附試驗等。簡單地說,在ELISA中,一個未知量的抗原附著在一個表面上,然后一個特定的抗體被施加在表面上,以便它可以綁定到抗原。這種抗體與酶相連,在zui后一步添加一種物質,這種酶可以轉化為一些可檢測到的信號,通常是化學底物的顏色變化。
elisa試劑盒的抉擇 請認準信帆生物
If a protein with multiple epitopes is being detected, a sandwich assay is a good choice. It usually requires two antibodies that react with different epitopes. However, if the molecule has multiple repeating epitopes, it is possible in a sandwich assay to use the same antibody for both capture and detection. Alternatively, if there is a supply of the analyte to be detected in pure form that can absorb effectively to a microwell, then one can set up a competitive assay in which the purified analyte is immobilized and analyte in the sample competes with the immobilized analyte for binding to labeled antibody. In this case it is essential to titrate the antibody so that it is limiting, or else the assay sensitivity will be lowered.
elisa試劑盒的抉擇 請認準信帆生物
Polystyrene will bind a wide variety of proteins in an increasing amount depending on their concentration in the coating solution. The specific and optimal amount needs to be determined for each protein, but some general observations have been made for antibodies. Medium to low binding plates bind typically up to 100 - 200 ng of IgG/cm2 while high binding plates typically can bind up to 400 - 500 ng of IgG/cm2. In addition to proteins, polystyrene plates will absorb peptides generally of 15 - 20 amino acids in length. In order to achieve strong binding, a peptide will need both hydrophobic and hydrophilic interactions. Typically a drawback to adsorbing peptides directly is that they tend to have few epitopes, and if these are involved in interaction with the plastic, it will be difficult for an antibody to bind to them. One alternative is to attach the peptide to a larger protein through a spacer arm that provides some distance between the peptide and the protein, allowing the antibody to interact with the peptide.
An organism such as bacterial or viral assays that detect whole organisms can also use sandwich assays with the same antibody for both capture and detection. If the target molecule is small or consists of a single epitope, a modification of the formats described above is needed. Small molecules by themselves either do not adsorb well to a solid phase, or may be masked by the blocking protein added. However, small molecules can often be attached to larger proteins which provide a means to attach the desired epitope to a solid phase in a configuration that allows the epitope to be bound by an antibody.
Carbohydrates and heavily glycosylated proteins do not adsorb well to polystyrene by the forces described above because they have very little ability to participate in hydrophobic interactions. Membrane proteins released from cells and maintained in solution by detergents are also not adsorbed well in the presence of detergents. Covalent linkage or reduction of the detergent concentration are the best means for attaching these proteins. In fact, covalent linkage can be performed in the presence of detergents such as Tween-20 and Triton X-100.