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| Observation | Problem | Corrective Action |
|---|
| No signals on positive control spots | Concentration of detection antibody and/or SA-HRP too low | Use concentration/dilution specified |
| Chemiluminescent reagents failed | Repeat with fresh chemiluminescent reagents according to manufacturers protocol |
| Inadequate exposure time to film | Increase exposure time |
| Observation | Problem | Corrective Action |
|---|
| No or low signals on target spots | Sample dilution too high | Use more sample |
| Analyte abundance in sample is low | Use more sample |
| Use different cell line |
| Verify that conditions used to stimulate cells were optimal |
| Sample deteriorated during preparation | Supplement buffers with protease and phosphatase inhibitors where indicated |
| Sample deteriorated during storage | All samples should be stored at ≤ -70 °C. |
| Avoid freeze-thaw cycles |
| Inadequate exposure time to film | Increase exposure time |
| Observation | Problem | Corrective Action |
|---|
| Signals on negative control spots | Concentration of detection antibody and/or SA-HRP too high | Use concentration/dilution specified |
| Sample concentration too high | Use less sample |
| Observation | Problem | Corrective Action |
|---|
| Uneven or high background on blank areas of array | Insufficient washing | Perform number of washes with volume indicated as specified in the product insert protocol |
| Perform washes in large container and not the 4-Well Multi-dish |
| Concentration of detection antibody and/or SA-HRP too high | Use concentration/dilution specified |
| Array was allowed to dry out partially during the procedure | Always keep arrays submerged. Minimize time the array is exposed to air. |
| Creased or scratched arrays | Use flat-tipped forceps to handle arrays on array number. |
| Observation | Problem | Corrective Action |
|---|
| Too many spots have signals | Insufficient washing | Perform washes as stated in product insert. Perform washes in a large container, not in the 4-Well Multi-dish. |
| Concentration of detection antibody and/or SA-HRP too high | Use concentration/dilution specified |
| Sample concentration too high | Use less sample |
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