4 replies to this topic

[rick112]

hi

we all get weird artifacts now and then in SDS-PAGE, strange vertical or horizontal lines, blotches, spots, etc..these are mainly seen when SDS-PAGE is developed with silver staining…
lets all pool in from our experience what are the common artifacts seen in SDS-PAGE and what are there reasons ???

[neuron]

I get horizontal lines when i use once used Tris glycine buffer

[K.B.]

In my experience vertical lines are most probably impurities in the sample or sample/lysis buffer.

[rick112]

K.B., on May 8 2009, 05:05 PM, said:

In my experience vertical lines are most probably impurities in the sample or sample/lysis buffer.

ya but they could also come if the "Comb" used are not cleaned properly....i also remember getting horizontal lines ones...and we found out finally that it was some problem with the water we used to make the running buffer...

some times i have seen is peculiar problem where a particular portion of the gel (lets say lower region or upper region) captures more stain than rest of the gel...any one ever faced this problem???

[mdfenko]

one of the common artifacts seen in sds-page when using reducing agent (especially with old reducing agent) comes from keratins from dust. you see horizontal bands (usually all the way across the gel) in the range of 55-68 kDa.

they are prominent when silver stained and when developing western blots with color deposition (ap, nbt, bcip).