Antibody selection requirements in immunohistochemistry Marketing Department Beijing Bayerdi Biotechnology Co., Ltd.
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Antibody selection requirements in immunohistochemistry
1, primary resistance selection points
(1) Select monoclonal or polyclonal antibodies. A specific antibody produced by a clone is called a monoclonal antibody, and a monoclonal antibody can be specifically targeted to a single specific antigenic determinant, just as a missile accurately hits a target. On the other hand, even with the same antigenic determinant, antibodies can be produced from several clones in the body, forming several monoclonal antibody hybrids called polyclonal antibodies. In the antigen-antibody reaction, the general monoclonal antibody is specific, but the affinity is relatively small, and the sensitivity of the detection antigen is relatively low; while the specificity of the polyclonal antibody is weak, the affinity of the antibody is strong, and the sensitivity is high, but non-specific staining is prone to occur ( Can be avoided by blocking etc.).
(2) Source of species. Generally, rabbit-derived antibodies are mostly polyclonal; while mouse-derived antibodies are mostly monoclonal, but there are also others. This article is mainly to match the source of the secondary antibody in the back.
(3) The purpose of the experiment is to detect what kind of antigen, ie species reactivity. This is very important, as indicated in the general instructions, such as mouse Ms, rat Rat, human Hum and so on.
(4) Can you do immunohistochemistry? Generally, the first-resistance instructions will be marked as WB, IHC, ICC, IF, etc. It is recommended to select the indicated antibodies, as they are generally proved by the article.
(5) Test specimen type. For the detection of paraffin sections or frozen sections, antibodies that can generally be used for paraffin sectioning may be used to detect frozen sections, but antibodies that can be frozen sections may not be able to detect antigens in paraffin sections.
(6) Manufacturer. The quality of the original antibody produced by famous foreign antibody manufacturers is generally no problem, but it is a bit expensive; and the primary anti-working liquid used by domestic distributors is cheap, but sometimes the stability and repeatability of the results are slightly worse, especially the repeatability of different batches. difference.
(7) The contradiction between price and quality. The quality of an antigen pack may be better because many primary antibodies avoid repeated freezing and thawing and are less stable in the diluent (if the time is long), but the price is more expensive.
2, secondary resistance selection points
(1) Source of species. The source of the secondary antibody is determined mainly according to the source of the primary antibody. For example, the primary antibody is the source of the mouse, and the secondary antibody can be purchased from the anti-mouse (the sheep, the rabbit, etc.).
(2) Selection of markers. There are markers such as HRP, Biotin, and fluorescein. Generally, the SP three-step secondary antibody selects the Biotin-labeled secondary antibody to bind to the latter SP; and immunofluorescence staining requires the purchase of different fluorescein-labeled secondary antibodies, such as rhodamine, FITC, Cy3 and the like. This is mainly related to the presence or absence of the third and third antibodies.
(3) Choose IgM or IgG. Generally, the primary antibody is IgM, then the secondary antibody selects IgM; otherwise, the primary antibody is IgG, and the secondary antibody selects IgG.