![]() However, tissue lysates and tissue culture supernatants that contain serum will contain immunoglobulins. An example is western blotting of a cell lysate that is not expected to contain IgG. A primary antibody raised in rabbit would be an appropriate choice, followed by an anti-rabbit IgG secondary antibody.įor techniques using samples that do not contain endogenous immunoglobulin (IgG), the choice of host species of the primary antibody is less critical. For instance, if you are studying a mouse protein, choose a primary antibody that is raised in a species other than mouse. This is to avoid cross-reactivity of the secondary anti-immunoglobulin antibody with endogenous immunoglobulins in the sample. The species the primary antibody is raised in should be different from the species of your sample. Choosing the species of primary antibody host A prediction of cross-reactivity is made based on sequence similarity. If your sample is not from one of the species listed in the datasheet, this means that the species has not been tested and we cannot demonstrate suitability. The antibody may react with the same target protein from other species sharing sufficient amino acid sequence homology. If possible, choose an antibody that has been raised against the same species your sample is from. These restrictions on use are noted in the applications section of the datasheets. Others cannot bind to their targets in formalin-fixed, paraffin-embedded tissues without an antigen retrieval step that reverses the cross-links introduced by formalin fixation. Our antibodies for western blotting require the samples to be reduced and denatured unless otherwise noted on the datasheet.įor immunohistochemistry, some antibodies are only appropriate for unfixed frozen tissue. On the other hand, some antibodies will only recognize epitopes on proteins in their native, folded state. Many antibodies will only recognize proteins that have been reduced and denatured, because this reveals epitopes that would otherwise be obscured by secondary and tertiary folding of the proteins. Some antibodies require samples to be treated in a specific manner. For example, if you are trying to detect a cell surface protein on live cells by FACS, choose an antibody that is raised against an extracellular domain of the protein. The immunogen is generally described on the datasheet (in some cases an exact description of the immunogen is not given for proprietary reasons).Ĭheck that the immunogen is identical to or contained within the region of the protein you are trying to detect. Immunogens can be full-length proteins, protein fragments, peptides, whole organisms (for example, bacteria), or cells. The region of the protein that you wish to detectĪntibodies are generated by immunizing host animals with an immunogenic substance. The nature of your sample will determine which antibody is most appropriate. For assistance with your application, browse our protocol library. Out antibodies are continuously tested in-house and datasheets are updated with latest application information. If an application is not listed, it means that we have not tested it and it is unknown how the antibody will perform. When an antibody is tested in an application and fails, this is also noted on the datasheet. Antibody datasheets list the applications we have successfully tested the antibody in. ![]()
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