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I recently ordered your Twinkle antibody (ab83329) and am having
some trouble identifying the proper band. I ran a 12% gel and
loaded 20ug of protein (isolated using RIPA buffer), probed with
1:1000 of Twinkle (overnight) and secondary rabbit at 1:1000 for
I also loaded the Jurkat cell lysate as the datasheet recommended
it as a positive control. I am needing help with identifying which
band is the correct one. The predicted size suggests 77kDa along
with an unknown band at 57kDa. The Jurkat cell lysate has a strong
band that falls between the 70kDa and 55kDa marker which suggests it
is the 57kDa unidentified band.
The first lane does appear to have a band between 70kDa and 100okDa,
which, suggests that it is the proper size. Do you think so? However,
if that is the case, why didn't the Jurkat lysate turn out
Asked on Jun 14 2012
Thank you for contacting us.
The human protein has 3 isoforms: 77, 66, and 60 kDa (http://www.uniprot.org/uniprot/Q96RR1).
Thus, the band you are seeing around 57 kDa could be the third isoform at about 60 kDa. Please keep in mind that the MW marker are not precise and a difference by 2 or 3 kDa is possible.
The bands could vary from sample type to sample type - depending on which isoform is present, and under which conditions the isoforms exist.
I assume you are using human samples?For another species, the results might vary as well.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
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Answered on Jun 14 2012