Technical Service Walkthrough
Antibody / antiserum / protein
Specify the name of the product:
Nature of the problem and previous experience
1. What is the specific problem you are experiencing?
2. Did the same vial of product work in the past? What were the results?
3. Did other lots of this product work in the past? Which lots? What were the results?
4. How have you reconstituted the lyophilized product (diluent, volume)?
Samples and controls
1. Species (animal) of the sample
2. Type of sample (organ, slice thickness, fixation procedure)
3. What positive and negative controls were used? What were the results?
Performing the experiment
1. Was a blocking performed? If yes, describe it (blocking solution, time, temperature).
2. At what temperature has the primary antibody been stored? Has it ever been frozen?
3. At what dilution was the primary antibody used and what was the diluent?
4. What were the incubation conditions (time, temperature) for the primary antibody?
5. Have you performed washing between the first and the secondary antibodies? Of what kind?
6. What was the secondary antibody used (company, HRP or AP conjugated, fluorescent-probes, made in which species, against which species, etc)?
7. At what dilution were the secondary antibody (or the avidin-biotin complex) used and what was the diluent?
8. What were the incubation conditions (time, temperature) for the secondary antibody (or the avidin-biotin complex)?
9. Has a secondary control been performed successfully?
10. What detection system was used for the enzymes?
11. Has dehydration been performed before covering of the slices? Which mounting media was used?
12. Would you have pictures to image the situation?
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