Single and multiplex immunolocalization protocols for protein and mRNA are available. Staff members have extensive experience with > 500 primary antibodies for animal and human fixed and frozen sections, cytospins, and cell cultures. Microscopes are available for photography.
Services
- Project planning, tissue selection, and experimental design including antibody selection.
- Testing antibodies or ISH probes
- Protocol development and validation
- Animal and human block archives for positive and negative controls
- Microscopy training or services.
Immunolocalization
- IHC: single and multiple (HRP, AP, OPAL/TSA)
- IF/Multiplex IF: secondary antibodies provided for a wide range of host species (rabbit, mouse, guinea pig, rat, goat, sheep).
- Leica Bond autostainer for batch staining up to 36 slides (3 sets of 12 slides)
User provides primary antibody and slides. A small selection of free primary antibodies is stocked.
For new antibody testing, please contact Dr. Ann Fu, Core Lab Manager, at 352-273-7749 or fudo@ufl.edu.
For initial testing of new antibodies, different antigen retrieval methods may be tested using a known positive sample before testing on your sample of interest. Serial dilutions of the antisera are done to achieve the most specific and sensitive titer. Negative controls include the following: substitution of primary antibody with host species IgG, antisera peptide pre-absorption, and/or experimental tissue controls.
When an antibody lot is changed or an antibody has not been used for several months/years, verification is performed by testing the last known set of validated conditions on similar samples. Once verified, experimental samples can be run without further validation. If antibody performance cannot be reproduced, the antibody may undergo revalidation or an alternative antibody could be chosen. These options along with other possible approaches can be discussed during the initial consultation.
Microscopy
Microscopes for brightfield, fluorescence, laser microdissection, and whole slide scans can be used for stained slide analyses. Assisted and self-service use is available.
RNA In situ hybridization (ISH/FISH)
RNA detection services are available with users providing probes and slides. ACD RNAScope probes are recommended. Assay optimization is performed by varying sample digestion temperatures and times.
“… Immunohistochemistry is technically complex, and no aspect of this complexity can be ignored, from the moment of collecting the specimen…”
– Taylor, CR., Arch Pathol Lab Med
Fees and Invoicing
Please visit the Fee page for information on costs and billing.