Cell Marque generally recommends the use of in vitro diagnostic (IVD) reagents for IHC (whether the final application is for IVD or RUO) since these are manufactured based on a validated process. Often, Research Use Only (RUO) antibodies offer no assurance of such validation and moreover such products may not even be manufactured in accordance with cGMP (Current Good Manufacturing Practices). Cell Marque, of course, always uses a validation process and follows cGMP and ISO 13485:2003 standards. Cell Marque will, on occasion, market RUO antibodies; however, these are still manufactured to the same rigorous standards as Cell Marque's IVD antibodies.|
It is also important to consider whether a marker has been published in peer reviewed journals; however, it may take years for such publications to appear and the pace of introduction of new markers has increased greatly in tandem with a greater understanding of the human genome. The question arises as to whether it is worth waiting years for such publications or whether a lab should perform its own validation and correlation studies, aiming to make use of the best tools as soon as possible. In this circumstance, many labs elect to make such determinations independently and to seek correlation with a retrospective study that indicates good sensitivity and specificity.
Also, keep in mind for each step of the validation, it is best to change only one variable at a time and keep all the other variables constant. For example, one should never try different pretreatments, titers, clones, and detection kits all at the same time. Doing so will obscure what promotes and what detracts from optimal staining and therefore will interfere with the goal of producing consistent, optimal staining.
When one changes any step in a standard protocol, one will also need to revalidate. If this means switching to the best markers or chemistry for a given application, such a choice is to be commended because it is a reflection of a laboratory's commitment to the highest standards of patient care.
Start by troubleshooting the problem yourself; Review and consider the troubleshooting factors:
- If you need assistance to troubleshoot, contact a trusted expert in order to brainstorm
- Try to implement suggestions; Re-run the procedure on an immunoreactive, positive control
- If you don't get staining, look for another positive control; Repeat the suggested protocol
- If not successful, send the manufacturer your positive control for evaluation and ask for their reccommended protocol
- If not satisfied with the manufacturer's recommendation, you may exchange or return the product