I have tried a few times and have gotten many bands- probably do to the various isoforms it has. Any suggestions? I read that you need to run a nondenatured gel. However, can you get away with just eliminating BME (beta mercaptoethanol)
What antibody are you using? Is there any info on the data sheet about the bands it gives under denaturing conditions? Do you want to quantify the amount of collagen in your skin tissue? In that case, you might be able to do a combined quantification of all the bands (if you sure they are just different isoforms?). Or try a different antibody? Unfortunately some antibodies just don't work. :(
Thanks- It does work on cells- fibroblasts- we get a mature and an immature band. But I can't seem to get it to work with tissue.
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