The gel picture attached is the outcome after performing SDS-PAGE . I dont know why only one band are seen for my protein??
gel picture is not clear , please up load some more details such as kind protein, percent of gel used , etc.
It was venom protein fraction(15 microgram). 12% SDS separating gel and stacking gel was used.
For separating gel: Water 3.35ml
Bis-acrylamid: 4.0ml
Tris HCL: 2.5 ml pH 8.8
10% SDS solution 100 microlitre
10% APS: 50 micro
TEMED: 5 micro
Stacking gel : Water 3.35ml
Bis-acrylamid: 4.0ml
Tris HCL: 2.5 ml pH 8.8
10% SDS solution 100 microlitre
10% APS: 50 micro
TEMED: 10 micro
You have really high background I suggest to use apropriate destaining buffer for a long time. However, the problem could be in separation procedure like too long time, too high voltage or breakdown of proteins to low molecular weight components which you observe in head band.
Anna Kononiuk Can you plz suggest prefect voltage and running time in SDS?
It is depend on what MW proteins you expect, for small gel usually 15-30V through stacking gel and then 60-100V.But if you have only low molecular weigh protein it doesn't help. Fristly try to destein that gel with solution of Et-OH: acetic acid: water (8:2:1).
I had prepared destaining solution according to ur instruction & solved the problem. Thank u so much for being helpful
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