I would like to ask if immunohistochemistry must use antibodies of the same species as laboratory an

No. Because there are different types and quantities of antibody receptor FCRs on the surface of the cells of the same animal, you may have a very strong non-specific signal when you choose an antibody from the same animal as the primary antibody.

If a certain antigenic molecule of human is detected, the mouse anti-human antibody can be selected as the primary antibody, and then the rabbit anti-mouse antibody can be used as the secondary antibody. The primary antibody selects the species that recognizes the animal you want to label, and generally does not produce the same species as the animal protein you want to label.

Choose the primary antibody that is specific to you.

For example, if you want to label a specific protein, first buy a primary antibody that can label the protein, the species is not limited, but it must be able to identify the protein of the species you want to label. If you want to label the rat actin protein, then buy an anti-actin primary antibody that recognizes rat actin (it may also recognize other animals, this doesn't matter), it is generally ** common animals such as rabbits, horses, sheep, etc.

If you want to ask whether anti-rat actin can be used to label mouse actin, this is not absolutely no, the antibody may have some cross-reactivity in different species. But no one can guarantee this effect.

Secondly, if you choose a primary antibody of the same species as the experimental animal, then the secondary antibody used to link the primary antibody must be IgG against the animal, then it will bind to the other proteins of the sample, because the secondary antibody is species-specific, so the background will be high.

If you want to buy a commercial antibody, you probably won't buy the same primary antibody you mentioned.

Immunofluorescence is also a tissue section, which should also be possible, compared with immunohistochemistry, but the secondary antibody is different.

For immunofluorescence, both monoclonal and polyclonal antibodies are acceptable, and monoclonal antibodies are generally preferred.

No, because the sections used in immunofluorescence and immunohistochemistry are generally different, one is a frozen section and the other is a paraffin section, so the degree of modification of the target protein on the section is different, so in many cases it cannot be universalized.

Immunofluorescence technology, also known as fluorescent antibody technology, is one of the earliest labeled immunotechnologies. It is a technique built on immunology, biochemistry, and microscopy techniques. For a long time, some scholars have tried to bind antibody molecules to some tracer substances, and use antigen-antibody reactions to localize antigenic substances in tissues or cells.

Immunohistochemistry is the application of the basic principle of immunology - antigen-antibody reaction, that is, the principle of specific binding of antigens and antibodies, through chemical reactions to make the chromogenic agents (fluorescein, enzymes, metal ions, isotopes) of labeled antibodies develop color to determine the antigens (peptides and proteins) in tissue cells, and to localize, qualitatively and quantitatively study them, which is called immunohistochemistry or immunocytochemistry ( immunocytochemistry)。

The sections used in immunofluorescence and immunohistochemistry are generally different, one is frozen and the other is paraffin, so the degree of modification of the target protein on the section is different, so in many cases it cannot be universalized. Qi's Biotech suggests that if they can be universal, which company's antibody instructions still distinguish between IF and IHC, wouldn't it be better to write them all! Therefore, when choosing an antibody, you must find the specific use you need in the use column.

Generally, the antibody manual has a detailed introduction, it is recommended that you read the manual carefully or consult the seller.

Dear, you can't do immunohistochemistry.

Personally, I think it should be possible, but it is recommended that you consult Qi's Biotech, which is more credible and reliable.

It's better to buy and write immunohistochemistry to do.

Otherwise, there is no signal.

or non-specific signals.

How do you know if it's an antibody problem or a problem in the middle?

And it's quite laborious to do an immunohistochemistry.

Just get things ready when you have to do it.

Protein is extracted by yourself, such as removing serum after administration and then preparing a sample of sincere protein, and antibodies are used to see the results of your action. What do you need to distinguish? I don't understand what you mean.

Yes, these two are conceptually a kind of technology, what you said is that cell crawling and tissue sections can use the same antibody, the principle is the same, but pay attention to the dilution of the primary antibody, as well as the section, pay attention to the fresh material fixation, to ensure that the protein in your tissue is not decomposed, otherwise, the antigen is not or denatured, and the antibody can not be recognized.

Antibodies commonly used in immunohistochemistry experiments are monoclonal and polyclonal antibodies. As far as I know, all that is used in tumor autologous immune cell technology are cytokines and tumor cell antigens.

The purpose of blocking with non-immune animal serum homologous to secondary antibodies in immunohistochemistry is to reduce non-specific binding. If you block the specimen with mouse serum, the serum may contain the protein of interest you are detecting, and then the primary antibody will bind to it and interfere with the results. And if you use sheep serum for blocking, it will not serve the purpose of blocking, that is, even if the blocking serum is combined with some non-specific sites, but because it is sheep, the secondary antibody will also bind to it, and then there will be a false positive result!

Therefore, it is best to block with normal animal serum of the same source as the secondary antibody.

One is enough, not in the number of experimental methods, but in whether the article is substantial, if the indicators are relatively substantial, such as studying receptors and ligands at the same time, or monitoring multiple upstream and downstream indicators in a signaling pathway at the same time, etc., it is not much of a problem to only do immunohistochemistry and invest in the domestic core. If the indicator is thin, you can add WB or PCR.