Frequently Asked Questions (FAQs)
I am planning to do IHC-related project. Where can I get help?
The Immunopathology Research and Development Lab, situated on 6th floor of the Veterinary Research Tower (Rm # T6-003), routinely provides assistance in IHC-related projects across the campus. The lab services include but are not limited to: developing and troubleshooting immunoperoxidase and immunofluorescence IHC, in situ hybridization, and TUNEL assay in a variety of species of fixed / frozen tissue, and cultured cells. The lab also provides services for preparation and cryosectioning of frozen tissues. Please email to Lynn Dong at email@example.com for any questions.
Prior to start IHC-related project, can I request a consultation meeting with Lynn Dong?
Absolutely. You’re welcome to schedule a consultation meeting with Lynn Dong at firstname.lastname@example.org. This is an effective way for me to understand your needs and to have you better informed with available service options.
I have trouble getting my target antibody to work properly. Do you provide troubleshooting?
Yes. Please email Lynn Dong at email@example.com to schedule a meeting to discuss the issue.
How many slides should I submit for a new antibody optimization?
Please submit six (6) slides of the positive tissue, with two (2) sections per slides (if possible), on positive charged slides. If the tissue is too big to fit 2 sections on a slide, please submit 12 slides for optimization.
How much antibody do you need for a validation?
The amount of antibody needed for successful IHC staining is dependent on an antibody titer and the density of your target protein in a particular tissue. To get a start, please bring me 20 ul of each of the antibodies to be tested.
What do I need to provide for antibody optimization/validation?
Please provide the following: a) the name of the PI, Dept info, account #; b) six (6) slides (see above); c) an aliquot of your target antibody (see above); and d) the spec sheet or data sheet or any background info for the target antibody.
What is your turn-around time?
The normal time is about 3 weeks, starting from the time of receiving of all materials to having the IHC results available for your review. It may vary depending on the amount of work received in any given time period.
Once an antibody is optimized by the Immunopathology R&D Lab, how many slides do I need to submit for each sample?
2~3 slides per sample. If you have more than 10 samples, please use ProbeOnTM Plus slides. As such, all samples can be processed in a batch to save reagents and minimize variations.
I have a research specific antibody, can you optimize/validate the antibody for me?
Yes. The Lab will provide antibody validation for any antibody of your interest.
Do I need to take training in order to use the facility?
Yes, you’re required to take training before you may use the lab facility. The training includes two parts: part 1 - introduction of various IHC options, theory, background, as well as planning and designing your particular IHC (~2 hrs); part 2 is hands-on IHC training (whole day). This training also covers future on-going trouble shootings with your IHCs. After training, you may reserve to use the lab facility to perform your IHC with your primary antibody, and the lab will supply all remaining reagents.
If an antibody worked previously, is it guaranteed to work all the time?
No. Fixation and other tissue processing steps can greatly affect the results of IHC staining. If you want to get consistent results for comparison, please standardize tissue processing procedure, especially pay attention to fixation step. For an example, “When is the tissue put in the formalin? How long has it been in the formalin, how big is the block, what’s the fixation type? How fresh is the formalin? What’s the pH? What’s the total time?” It becomes increasingly evident that two sections from one sample can be diagnosed very differently, due to the differences in sample preparation and fixation.