There are four areas to consider in antibody-specific selection

Antibody immune cells secrete immune substances, is the core of the mammalian adaptive immune system against pathogens, it is stimulated by pathogen stimulation by plasma cells (effector B cells) secreted by an immunoglobulin, can recognize and bind to specific pathogen antigens, and then by mediating neutralization, conditioning, effector T cell killing to clear the pathogen
.

With the rapid development of antibody preparation and modification technology, antibodies have been widely used in various fields of life sciences, including scientific research, diagnosis, treatment and so on
.

The essential meaning of specificity is "Connected with one particular thing only", that is, it is only related to a specific thing that is only yi, with specificity
.

In immunology, we will say "antibody is specific", which means that the antibody is specific, and a specific antibody can only bind
to only one antigen.

In fact, it is a class of antigens with specific antigenic epitopes, because some different antigens have the same antigenic epitope (called cross-antigens) and can also bind
specifically to the same antibody.

Scientific research work is inseparable from antibodies, WB, IP, IHC, IF, ChIP and FC all need to use antibodies, these experimental methods in the specific selection of antibodies need to consider the main points to understand, the specific selection mainly needs to consider four aspects: protein specificity, species specificity, experimental method specificity, marker specificity
.

1.
Protein specificity for the protein to be detected to find antibodies, several details to distinguish: a.
recombinant protein is not full-length expression, you need to pay attention to whether the immunogen region of the antibody is in the recombinant protein region
.

b.
Endogenous proteins can be clear about the way they are sheared and modified, and proteins of special phenotypes need to be sequenced and combined with antibody immunogen sequences to view
cross-reactions.

c.
Phosphorylated protein detection needs to determine specific sites, and phosphorylation at different sites means that there may be different mechanisms, which should not be generalized
.

2.
Species-specific isologous proteins vary greatly or slightly in
different species.

a.
At present, most of the commercialized antibodies are recombinant proteins or design polypeptide antigens based on human protein sequences, and cross-react with other species according to the homologity of proteins
.

Information on reactive species indicated in the instructions is required
.

b.
Some rare species are difficult to find antibodies, can be compared by the sequence of proteins, choose homologous sequence immune antibodies, but generally in this case, manufacturers will not accept their complaints about quality, if you can apply for free antibody samples, it is conducive to antibody selection
.

3.
Experimental method specificity At present, there are many experimental methods for using antibodies, different use methods in the process, because the treatment of protein samples is not the same, the content of proteins is different, and the identification epitope and titer requirements of antibodies are different
.

4.
The specificity of the marker is generally based on the experimental operation of the experimental method will not use the labeled primary antibody, such as WB, IHC, etc.
, are through the secondary antibody class of reagent labeling to achieve the purpose
of the results.

However, some experiments based on instrumental analysis may use directly labeled primary antibodies, such as flow cytometry
.

Then you need to know the fluorescence range that the instrument you will use can detect, and select the corresponding marker for the parameter requirements that are not comprehensible
.

In immunofluorescence double-standard experiments, different fluorescent markers
need to be selected.