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Fig. 2. Purification steps for the purification of recombinant laminin-1
α
1 G domain.
(A) Recombinant G domain (
) elutes within or as a shoulder off the main protein
peak (
) of a heparin-sepharose column. Molar concentrations of the NaCl eluant are
indicated (
). (B) Although recombinant G domain remains in the unbound fraction
of the DEAE-Sephacel column, it removes other unwanted proteins. (C) Elution profile
for recombinant G domain off of a heparin-5PW HPLC column. Pooled fractions
36-44 contain recombinant G domain whose purity is greater than 90%. Bars in each
panel represent fractions pooled at each stage of purification.
suspension, as opposed to monolayers, greatly increases cell numbers that will
reflect greater levels of recombinant produced. However, growth at 27
C is also
optimal for yeast and fungi, so careful aseptic techniques should be used at all
times.
°
3.
The pVL1392/pVL1393 series of baculovirus expression vectors has been excel-
lent for production of recombinant laminin domains in Sf9 cells in our labora-
tory, provided an appropriate signal sequence has been cloned in the vector. The
fibronectin signal sequence has successfully used to obtain secretion of a number
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