There are same thick bands of unknown protein. I'm not sure whether this band because of containing of salt or because of the low of molecular weight. I need some discussion or sharing experience from you. Thanks a lot ..
You must run marker with your proteins samples.
The judge may be done only on the base of marker appearance.
Also, your gel has not good resolution and quality.
This may be due to interferring materials in the samples such as ions and ...
I agree that the resolution of your gel is not very good. Usually, precast gels have a stacking layer ontop of the resolving layer. To me it looks as if the power supply was run at 200V and the samples did not stack prior to entering the resolving layer. I usually run the gel at 75V until the samples form a thin line right above the resolving layer then increase the voltage to 200 for resolution. I also do not use my SDS-PAGE running buffer more than 5 times.
The presence of dye in the wells suggests that not everything is entering the gel. Was BME added to the sample loading dye? Were the samples heated at 95C prior to loading? It looks to me that these samples derive from a crude lysis of organisms and cellular debris is messing up the SDS-PAGE run.
The other responders make good points. If you provide more information about the gel, running conditions, and size of product you were looking to resolve it would help solve your issue.
thank for the comments :). I run the protein from the plant sourse, ( i isolated the protein). The sample heated at 95 C before loading .The condition were : 12.5 % of polyacrylamide, running condition : 180 V, 400 mA, loading volume was 20 microlitre.
Your current is too high for this PAGE and can warm your gel.
Please keep your gel cool by cold buffer and placing running set in ice chamber
This help to finding better gel
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