Immunohistochemistry Assurances

 

No guarantees can be made for the successful application of immunohistochemistry (IHC) using new or novel antibodies. The quality of IHC is dependent on the quality of the antibody (Ab) and tissue. However, several assurances can be given based on experience and the use of appropriate controls during novel assay development. If a validated antibody is available, a particular assay can be reproduced with a minimal amount of effort. We typically include one of our tested 1° antibodies and tissue with your tissue of interest and your 1 antibody to ensure that our reagents are working. If we have a 1° Ab that labels your tissue of interest, we will also verify the quality of your tissue. All costs for these preliminary steps will be the obligation of the investigators. For a novel antibody we typically use multiple concentrations of the 1°and 2° Abs and a variety of blocking steps. From an initial experiment significant information can be obtained which can be used to guide subsequent improvements in the assay.

Investigators will have the ability to review tissue from the first experiment and discuss the results. Depending on the outcome, the investigator might choose another series of testing conditions or proceed to analysis of their tissue and conditions of interest. A second test procedure is usually recommended to optimize and verify the quality of the original staining. If the initial test was not satisfactory a variety of modifications can be suggested. However, the investigator will have the option of choosing not to proceed with that particular antibody.

Molecular Histology Center Environmental and Occupational Health Sciences Institute, 170 Frelinghuysen Road, Piscataway, NJ 08854 Phone: 732-445-3729 For additional information contact ekr@eohsi.rutgers.edu

Updated on Friday, June 03, 2005