I attached the zymogram image. The top line is MMP-9 giving distinct band. But the MMP2 band is like a straight line across the gel both latent and active forms even in the marker lane. Could any one suggest possible reasons for this pattern of zymogram.?
This may be an artifact of the loading buffer.. Is this the complete image ? Or just the top portions?
1.- Those are artefacts
2.- What's the molecular size of the upper bands? they look more like MMP2
3.- Can you post the complete image?
I think it is artifact. Is there any leaking from the well when you load your sample? Both MMP9 and MMP2 should display a clear band. I attached an image of MMP9 and MMP2 with zymography. Hope it is helpful.
Thank you for the reply. I guess it was because of leaking of sample while loading. Now I am getting the bands as you shown. Even though I am curious to know the reason for streaks. (I can see the same streaks below the MMP-2 lane in the gel you posted also) What do you suggest it was? I read a paper (http://www.ncbi.nlm.nih.gov/pubmed/11826121) showing the degradation of MMPs upon heating. But we are not exposing to more than 37 degrees. How it is coming below 55kd?
What I am thinking is two possible reasons: 1. sample leaking 2. votage too much high. Sometims, If you chage votage, it will give too much pressure on sample moving. Also, higer votage may increase the local temperature. I ma not sure how much it could change, but if MMP is sensitive to temperature, then it could be a possible reason.
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