EPITOMICS - IHC/ICC Troubleshooting Form
Please answer the following questions to help us troubleshoot your issue.
1. General Product Information
a. Product Name:
b. Catalog #:
c. Lot#:
2. Is this your first time using this antibody?
a. Yes, this is the first time
b. No, this is a re-use of the same tube of antibody
c. No, this is a re-order of a previously used antibody
3. Please briefly describe the issue that you are experiencing.
A:
4. How is the product currently being stored?
A:
5. Please describe the sample(s) used with the antibody.
Sample Type:
Sample species:
Tissue or Cell:
If Tissue: Is it fresh, frozen, or paraffin?
Type of Fixation (e.g. Methanol, Acetone):
% of Fixation:
Fixation time and temperature (e.g. overnight at 4C):
6. Was the sample deparaffinized? If yes, please briefly describe the method used.
A:
7. Was any pretreatment done on the sample? If yes, please briefly describe the pretreatment reagent and method used (e.g. 10mM Citrate Buffer in 20 minutes in Microwave, 98C)
A:
8. Primary antibody step. Please describe:
Dilution used:
Type of dilution buffer used:
Length of incubation with primary:
Temperature of incubation:
Volume of primary antibody used (µl)?
9. What type of wash buffer was used and how often/how long?
(e.g. PBS, twice, 5 minutes each)
A:
10. What blocking buffer/substrate was used, how long, at what temperature?
(e.g. 10% normal goat serum, overnight at 4C)
A:
11. Secondary antibody step. Please describe:
Type of secondary (i.e. goat-anti-Rb-HRP, Polymer, goat-anti-Rb-FITC) and supplier:
Length of incubation:
Temperature of incubation:
Dilution used:
Type of dilution buffer used:
12. Tertiary component/substrate step. Please describe :
Type of tertiary (i.e. SA-HRP) and supplier:
Length of incubation:
Temperature of incubation:
Dilution used:
Type of dilution buffer used:
13. For IHC, please describe the Chromagen and Counterstain used:
Type of Chromagen (e.g. DAB):
Length of incubation:
Temperature of incubation:
Dilution used:
Type of Counterstain (e.g. hematoxylin):
Length of incubation:
14. For ICC, please describe the Counterstain and mounting condition used:
Type of Counterstain:
Mounting solution used:
Was the slides kept in the dark after mounting?
15. Was a positive and/or negative control used? If yes, what were their results?
A:
16. Please include any other information that you wish to share.
A: