Fig. 3. CrkRS has an apparent Mr of 180 kDa on SDS-PAGE. (A)
Affinity-purified anti-CrkRS antibodies are specific for CrkRS. Immunoblot of
the GST-CrkRS C-terminal fragment (GST-CCT) fusion protein purified from
E. coli and probed with affinity purified anti-CrkRS antibodies. The
antibodies did not recognise the lane containing only GST (i), and
pre-clearing the anti-CrkRS antiserum abolished the CrkRS signal (ii). (B)
(His)6-tagged human CRKRS expressed in Sf9 cells runs at 180 kDa.
Anti-CrkRS immunoblot of lysates from Sf9 cells either uninfected, or infected
with wild-type, or CrkRS-expressing baculovirus. (C) In vitro
transcription/translation product of the CrkRS cDNA has an apparent
Mr of 180 kDa. Full-length CrkRS was transcribed and
translated in vitro from an expression vector containing the full length CrkRS
cDNA (lane 2). An empty expression vector was used as a control (lane 1).
