Product FAQs
1.Why are the bands blurred?

· Too high voltage or too long running time produces too much heat which increases temperature of the buffer. Please refer to product manual.

· Buffer is old. Use fresh solutions and pre-chill the buffer before use to get a great result.

· Gel concentration is low. Try proper gel concentration.

· Gel is old. Use newly made gels.

2.Why do bands become skewed or distorted sometimes?

· It is suggested to load protein marker to the middle lane because bands at the edges of the gel may become wider or distorted due to edge effects or ionic strength difference.

· The gel is uneven or contains bubbles. It is also possible that some gel residues exist in the sample well.

3.Protein Markers cannot be separated during electrophoresis.

 Mostly, gel concentration is not suitable, which should be within recommended concentration range. Sometimes buffer and gel are not fresh enough to lead to a good electrophoresis result.

4.The band of prestained protein marker is clear after electrophoresis, but become faint after protein transfer.

· The transfer sandwich should be prepared properly. Otherwise bands may become missed or faint in protein transfer.

· Transfer condition needs to be optimized. 200 mA of 2 hours or 100 mA overnight is recommended.

5.What is the methanol concentration during protein transfer? How it affects protein transfer?

The recommended methanol concentration is 15%. Too high concentration may cause protein passing through the membrane and finally transferred to the filter paper.

6.Is it necessary for ECL mix to be prepared in dark?

There is no need to prepare ECL mix in dark, but it is necessary to use fresh ECL mix and continue the following steps in the dark room immediately after ECL mix is prepared well.

7.How long is the optimal exposure time for ECL mix?

Light emission can last up to 2 hours. It is more intense during the first 30 minutes but decreases with time. Longer exposure times may be necessary as time elapses.

8.Why the background is black after exposure to X-ray film?

Exposure time or developing time is too long.

9.Why are there many unexpected bands for Western Markers after exposure to X-ray film?

It is due to too much sample loaded or too long exposure time. It is also possibly because of too high concentration, low specificity or low purity of secondary antibodies.

Message