6.1 No Signal

Cause Investigation

1. Sample-related Causes
(1) Sample does not express the target protein or has low expression
Select an appropriate sample and conduct proper stimulation treatment.
(2) Improper sample preparation
Add protease inhibitors and phosphatase inhibitors; pay attention to the details of sample preparation and the storage conditions after preparation to prevent protein degradation; select the appropriate cell components for sample preparation.
(3) Insufficient sample loading amount
Increase the sample loading amount. It is recommended to load 20-50 μg of protein for cell samples and 50-100 μg for tissue samples.
2. Antibody-related Causes
(1) Improper antibody selection
Select a suitable antibody, considering factors such as species reactivity and application scope.
(2) Improper antibody storage
Store the antibody in accordance with the instructions provided with the antibody.
(3) Insufficient antibody incubation time or concentration
Extend the antibody incubation time and increase the antibody incubation concentration.
3. Experimental Operation-related Causes
(1) Improper membrane transfer
Select a membrane appropriate for the molecular weight of the target protein; optimize the membrane transfer voltage, current, and duration. After membrane transfer, use Ponceau S or Coomassie brilliant blue stain to verify the membrane transfer efficiency.