S:\SOP Office\Marketing Protocols\TP102 - WB Troubleshooting.docx
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TP102–WB Troubleshooting
In order for us to help you better and faster, please provide us with as much information as possible by filling out this form and returning to Abbiotec Technical Support Department by email to or fax to 1 (858) 586-6252. Attach any relevant experimental data (positive or negative) with legend.
- General information
- Customer name:
- Catalog Number:
- Lot Number:
- Date received:
- Storage temperature:
- Was the product aliquoted?
- Lysate information
- Species:
- Cell Line:
- Primary Cells:
- Whole Tissue:
- Lysis buffer used:
- Protease inhibitors used in lysis buffer:
- Were the samples sonicated?
- How long the lysate was stored?
- Lysate storage temperature:
- Gel information
- Reducing or non-reducing gel:
- Supplier of the gel:
- Gel percentage :
- Sample running information
- Samples heating temperature and time:
- Amount of lysate loaded per well (ug):
- Was the running buffer re-used?
- Transfer information
- Type of membrane was used:
- Membrane transfer time:
- Membrane transfer current:
- Was the transfer buffer re-used?
- Was the blot stripped and re-probed?
- Blocking information
- Blocking buffer used:
- Blocking time:
- Blocking temperature:
- Primary antibody information
- Primary antibody dilution:
- Primary antibody dilution buffer:
- Incubation time for primary antibody:
- Incubation temperature for primary antibody:
- Washing after primary antibody incubation:
- Washing buffer:
- How many times was washed/how long:
- Secondary antibody information
- Secondary antibody supplier:
- Secondary antibody species reactivity:
- Secondary antibody dilution:
- Secondary antibody dilution buffer:
- Incubation time for the secondary antibody:
- Incubation temperature for secondary antibody:
- Washing after secondary antibody incubation:
- Washing buffer:
- How many times was washed/how long:
- Signal detection information
- Detection system used:
- Supplier of the detection system:
- Length of time for film exposure:
- Results
- What signal can be seen on the developed film?
- Bands (molecular weight markers, no bands or extra bands, wrong size, etc):
- How many times this Western blot was tried?
- Are the results the same every time?
- Was this antibody previously working on these samples?
- Positive and negative controls used:
- Different dilutions tested for primary antibody:
- Was a secondary antibody control (without primary antibody) run?
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