Help WB development is over,Why is the background so dark, it's so ugly!
In fact, in response to this problem of black background, you can check yourselfIs it that the primary antibody is not working well, or whether you have reused the primary antibody?At this time, it is recommended that you can use a new primary antibody, 4 incubate overnight to try
As one of the basic experiments, in addition to the above problem, we believe that we have also encountered such a problem:
High Background:The high background is likely to be due to poor blocking, high concentration of primary antibody, or the time and number of times you wash the membrane are not enoughIncrease the blocking time, decrease the concentration of the primary antibody, and increase the time and number of washes to solve.
Blank Sheet:There are many reasons for this, and the most common cause may be youThe primary antibody was added incorrectly, so check it out
The strip has a white circle with a regular edge:If the strip has a white circle with a regular edge, it is likely that there is a bubble between the film and the glue you are using, so you want itTake care to remove air bubbles between the film and the glue before transferring.
Strip trailing:This problem can be caused by too much protein, too high a concentration of the primary antibody, and too long an incubation time, depending on your experimental needsAdjust the amount of protein loaded, adjust the concentration and time of the primary antibody