Immunostaining

TACMASR offers immunohistochemical staining on formalin fixed, paraffin embedded (FFPE) tissues and cell lines, and on frozen tissues and cultured cells. Chromogenic techniques are available now and our future plans include fluorescence techniques (if there is sufficient demand).

Immunohistochemistry (IHC)

Ventana Medical Systems - Discovery XT instrumentTACMASR performs chromogenic immunohistochemistry IHC, on the Roche/Ventana automated staining platforms. We use validated reagents from Roche/ Ventana Medical Systems  for antigen retrieval and staining. Our instruments can stain FFPE, frozen tissue and cell pellets. They are also capable of performing In Situ Hybridization (ISH).

Primary antibodies used for IHC

TACMASR has a list of more than 80 commonly requested, validated antibodies that can be run on our instruments. We have validated well over a hundred antibodies for investigators, however some of them are not commercially available. Please contact our specialist, Ed Abril, to discuss your research needs.

Microscope image of immunohistochemistry - Rimsza lab

Antigen retrieval validation is used to optimizing conditions for the antigen of interest from formalin-fixed, paraffin-embedded (FFPE), frozen tissues and cell lines. Optimized conditions may differ depending on the antibody being used. The use of an automated staining platform allows for both run-to-run and intra-run reproducibility for each assay. The selection of primary antibodies we have validated to date is investigator driven, and we are continually adding new antibodies upon investigator request. Primary antibodies can be purchased or acquired from a number of vendors and sources. Please consult Ed Abril for the best commercial sources to use. 

We only stock those primary antibodies that are in high demand, but investigators are encouraged to purchase their own primary antibodies* that we have not yet validated. All other bulk reagents and IHC kits are supplied as part of the IHC assay.

A critical aspect of interpreting immunohistochemical and other assays offered by the service is the identification and use of the appropriate positive control tissues for each assay. TACMASR works with each investigator to identify and acquire appropriate pathological material and controls for IHC studies.

* If you are looking for candidate primary antibodies for IHC, check the vendor datasheet. Antibodies that provide information and images about it's use for IHC performed on FFPE tissues work best in our experience . We have found that antibodies that have only been validated against Westerns have a very low success rate (less than 25%) when used to stain FFPE tissue. We encourage users to read the 2015 NATURE news feature "Blame it on Antibodies", which discusses a number of issues related to antibody quality and reproducibility.