Hi Subhas
if all the lanes including the marker are problematic...there can be problem with the gel polymerisation....have you stained a section of that gel with Commassie.......
and 'not clear' means what...are they haizy, or diffused or too much background...?
If you can upload the image...it would be more informative to actually see the problem you are facing
bye
Dear Ajay
Thank u for your information
I have not tried with commassie staining....i will definitely stain with commassie.
I am attaching the gel image please have a look and suggest me for further modification.
Lane: 3 marker
Lane 2 Sample A
Lane: 1 Sample B
The resolution does of the gel is not proper, which could have happened for many reason:
1. Gel has not polymerized properly (try using fresh acrylamide-bisacrylamide, APS, etc).
2. pH of the buffers are not correct.
3. error in voltage during run.
I also faced similar problem few weeks ago and changing the acrylamide-bis stock worked.
good luck
Dear subhas,
1) the haizy bands in both marker and sample is mainly due to your resolving buffer pH problem or yr voltage fluctuation. First you check the pH of your separating and stacking gel buffer. the another thing is prepare your APS always fresh. n during your run just check the constant voltage is coming or not.
2) If you are not facing this problem in commassie staining then it may be due to the procedure which you are using for silver staining. Is it proper method u follow? because u didn't write here to method of silver staining.
if u want full protocol of silver staining just tell me i can give u.
Dear Maruf,
Commassie briliant blue is cheap, easy and best technique for protein staining, but if your protein concentration is too much lower then u can prefer silver staining protocol.
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