Immunohistochemistry (IHC) Staining
- Antibody stains are used to detect the presence of specific biomolecules (antigens) in tissue sections by the application of an antibody to that antigen.
- Provides unique staining to a given protein.
NSALabs® offers high quality immunohistochemistry staining services. Representative sample images of antibody stains performed by NSA are shown in our antibody gallery (click to view). NSA’s protocol for trials of new antibodies with our designated dilution series continually broadens the offerings we are able to make, including “in-house generated” antibodies.
New Antibody Trials/Titrations
NSA performs trials in small 12-well culture plates, using nine of the wells when performing our dilution series. The dilution series covers 8 dilutions plus one with no antibody. The dilution range is approximately in steps of one third from 1:150 to 500,000. That is: 1:150, 500, 1500, 5000, 15000, 50,000, 150,000, 500,000. NSA uses 2-4 ml per well of antibody solution. NSA’s vehicle solution is only TBS, with no BSA or other proteinaceous component. The lower viscosity of the solution is kinder to the sections. This follows the practice of Gloria Hoffman of University of Maryland, Baltimore. Once we establish the dilution, we process 10-20 large MultiBrain® sections per crystallizing dish (similar to a very deep petri dish), holding 60ml of solution. For previous and subsequent steps we use transfer baskets inside the crystallizing dish to minimize direct handling of the sections, thereby preventing tearing. Many of these implements can be found under NSA’s HistoTools®.
The antibody gallery is only a representative sample of antibodies we have performed. New antibody stains are performed weekly. If you do not see what you are looking for, please contact us or request a quote stating the antibody of interest.
Please note: Certain antibody stains require specialized protocols for perfusion and preparation of tissue. Consult with us prior to perfusion so that we may advise you on optimal handling. Contact us via our technical support form or call 865-675-2245.